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Genetic engineering applications in animal breeding
Hugo H. Montaldo
Abbreviations: ES: embryonic stem cells ESR: estrogen receptor locus IGF-I: insulin-like growth factor I MAS: Marker-assisted selection QTL: quantitative trait loci This paper discusses the use of genetic engineering applications in animal breeding, including a description of the methods, their potential and current uses and ethical issues. Genetic engineering is the name of a group of techniques used to identify,
Dis\\/Integrating Animals: Ethical Dimensions of the Genetic Engineering of Animals for Human Consumption
Traci Warkentin
\\u000a Research at the intersections of feminism, biology and philosophy provides dynamic starting grounds for this discussion of\\u000a genetic technologies and animals. With a focus on animal bodies, I examine moral implications of the genetic engineering of\\u000a ōdomesticatedö animalsŚprimarily pigs and chickensŚfor the purposes of human consumption. Concepts of natural and artificial,\\u000a contamination and purity, integrity and fragmentation and mind and
Dis\\/integrating animals: ethical dimensions of the genetic engineering of animals for human consumption
Traci Warkentin
Research at the intersections of feminism, biology and philosophy provides dynamic starting grounds for this discussion of\\u000a genetic technologies and animals. With a focus on animal bodies, I will examine moral implications of the genetic engineering\\u000a of ōdomesticatedö animalsŚprimarily pigs and chickensŚfor the purposes of human consumption. Concepts of natural and artificial,\\u000a contamination and purity, integrity and fragmentation and mind
Improving human and animal health using genetically engineered goats expressing lysozyme in their milk
J. D. Murray; E. A. Maga
The application of genetic engineering should not be undertaken lightly as it requires extensive infrastructure and inputs before the genetically engineered animal enters a breeding and selection scheme; it does not provide a mechanism for bypassing good animal breeding and selection practices. However, there are instances where GE can provide an opportunity to address a problem in animal agriculture for
NSDL National Science Digital Library
This remarkable Web site contains descriptions and animations of nineteen different kinds of engines. Engine types include internal combustion, steam, and sterling engines, and each page shows how the piston, crankshaft, and other components move together to generate power. The animations demonstrate the processes of intake, compression, and exhaust. Some of the featured engines have more detailed descriptions than others, and oftentimes, a brief account of the engine's history is included. One engine dates back to the early 1700s.
NSDL National Science Digital Library
Keveney, Matt
This website includes a variety of animations explaining the mechanical workings of a variety of steam, Stirling and internal combustion engines. The animations may be paused, slowed or sped up. The animations are accompanied by additional text explaining how each engine works.
Role of stem cells in large animal genetic engineering in the TALENs-CRISPR era.
Park, Ki-Eun; Telugu, Bhanu Prakash V L
The establishment of embryonic stem cells (ESCs) and gene targeting technologies in mice has revolutionised the field of genetics. The relative ease with which genes can be knocked out, and exogenous sequences introduced, has allowed the mouse to become the prime model for deciphering the genetic code. Not surprisingly, the lack of authentic ESCs has hampered the livestock genetics field and has forced animal scientists into adapting alternative technologies for genetic engineering. The recent discovery of the creation of induced pluripotent stem cells (iPSCs) by upregulation of a handful of reprogramming genes has offered renewed enthusiasm to animal geneticists. However, much like ESCs, establishing authentic iPSCs from the domestic animals is still beset with problems, including (but not limited to) the persistent expression of reprogramming genes and the lack of proven potential for differentiation into target cell types both in vitro and in vivo. Site-specific nucleases comprised of zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regulated interspaced short palindromic repeats (CRISPRs) emerged as powerful genetic tools for precisely editing the genome, usurping the need for ESC-based genetic modifications even in the mouse. In this article, in the aftermath of these powerful genome editing technologies, the role of pluripotent stem cells in livestock genetics is discussed. PMID:24305178
A Review and Rationale for the Use of Genetically Engineered Animals in the Study of Traumatic Brain Injury
Luca Longhi; Kathryn E. Saatman; Ramesh Raghupathi; Helmut L. Laurer; Philipp M. Lenzlinger; Peter Riess; Edmund Neugebauer; John Q. Trojanowski; Virginia M.-Y. Lee; M. Sean Grady; David I. Graham; Tracy K. McIntosh
The mechanisms underlying secondary cell death after traumatic brain injury (TBI) are poorly understood. Animal models of TBI recapitulate many clinical and pathologic aspects of human head injury, and the development of genetically engineered animals has offered the opportunity to investigate the specific molecular and cellular mechanisms associated with cell dysfunction and death after TBI, allowing for the evaluation of
NSDL National Science Digital Library
Students are introduced to the classification of animals and animal interactions. Students also learn why engineers need to know about animals and how they use that knowledge to design technologies that help other animals and/or humans. This lesson is part of a series of six lessons in which students use their growing understanding of various environments and the engineering design process, to design and create their own model biodome ecosystems.
Genetic Engineering & Xenotransplantation
NSDL National Science Digital Library
Shane Grey (Garvan Institute of Medical Research, Sydney Australia; )
The issue-focused, peer-reviewed article about xenotransplantation, or transplanting organs/tissues from other species to humans, offers hope to Type I diabetes sufferers because: insulin-producing tissue in animals (islets) has been isolated in tests, some human trials were successful with transplanted islets from human cadavers, and genetic engineering could create a 'super' islet that will survive after xenotransplantation.
Genetically engineered, live attenuated vaccines for Venezuelan equine encephalitis: testing in animal models
William D Pratt; Nancy L Davis; Robert E Johnston; Jonathan F Smith
The central objective of this research was to test molecularly defined, live attenuated Venezuelan equine encephalitis virus (VEEV) vaccine candidates that were produced through precise genetic manipulation of rationally selected viral nucleotide sequences. Molecular clones of vaccine candidates were constructed by inserting either three independently attenuating mutations or a PE2 cleavage-signal mutation with a second-site resuscitating mutation into full-length cDNA
Genetically engineered, live attenuated vaccines for Venezuelan equine encephalitis: testing in animal models.
Pratt, William D; Davis, Nancy L; Johnston, Robert E; Smith, Jonathan F
The central objective of this research was to test molecularly defined, live attenuated Venezuelan equine encephalitis virus (VEEV) vaccine candidates that were produced through precise genetic manipulation of rationally selected viral nucleotide sequences. Molecular clones of vaccine candidates were constructed by inserting either three independently attenuating mutations or a PE2 cleavage-signal mutation with a second-site resuscitating mutation into full-length cDNA clones. Vaccine candidate viruses were recovered through DNA transcription and RNA transfection of cultured cells, and assessed in rodent and non-human primate models. Based on results from this assessment, one of the PE2 cleavage-signal mutants, V3526, was determined to be the best vaccine candidate for further evaluation for human use. PMID:12922119
Rigby, P.
This series reviews new developments in recombinant DNA technology and its applications. Each volume consists of 3 - 4 mini-reviews. Volume 7 contains two articles on aspects of molecular parasitology and one review on gene expression in animal cells of biotechnological interest.
J. F. Kreuze; J. P. T. Valkonen; M. Ghislain
Genetic transformation through direct gene transfer methods holds promise for introducing novel traits to sweetpotato in cases\\u000a where no solutions by conventional breeding are available. This may be if the trait is not known in sweetpotato or it is governed\\u000a by complex inheritance. Sweetpotato is clonally propagated, highly heterozygous, polyploid and out-crossing ¢ in other words,\\u000a a challenging crop to
Myocardial and cerebral perfusion studies in animal models S65 In-vivo phenotyping of genetically engineered mouse models
Jegelka, Stefanie
of genetically engineered mouse models for amyotrophic lateral sclerosis is established by combining BT temper- ature (PC-SAM,SA Instruments,UK). MR experiments were performed on a 7T/8cm MR system (MRRS vasodilatation, rendering cerebral blood vessels more susceptible to vasoconstriction and reducing overall blood
Genetic engineering in floriculture
Yoshikazu Tanaka; Yukihisa Katsumoto; Filippa Brugliera; John Mason
The global flower industry thrives on novelty. Genetic engineering is providing a valuable means of expanding the floriculture gene pool so promoting the generation of new commercial varieties. Commercialisation of genetically engineered flowers is currently confined to novel coloured carnations. However, further products are expected given the level of activity in the field. In general terms engineered traits are valuable
Engineering visualization utilizing advanced animation
NASA Technical Reports Server (NTRS)
Sabionski, Gunter R.; Robinson, Thomas L., Jr.
Engineering visualization is the use of computer graphics to depict engineering analysis and simulation in visual form from project planning through documentation. Graphics displays let engineers see data represented dynamically which permits the quick evaluation of results. The current state of graphics hardware and software generally allows the creation of two types of 3D graphics. The use of animated video as an engineering visualization tool is presented. The engineering, animation, and videography aspects of animated video production are each discussed. Specific issues include the integration of staffing expertise, hardware, software, and the various production processes. A detailed explanation of the animation process reveals the capabilities of this unique engineering visualization method. Automation of animation and video production processes are covered and future directions are proposed.
Genetic aspects of animal reasoning.
Poletaeva, I I; Popova, N V; Romanova, L G
This paper reviews the investigations of Prof. L. V. Krushinsky and his colleagues into the genetics of complex behaviors in mammals. The ability of animals to extrapolate the direction of a food stimulus movement was investigated in wild and domesticated foxes (including different fur-color mutants), wild brown rats, and laboratory rats and mice. Wild animals (raised in the laboratory) were shown to be superior to their respective domesticated forms on performance of the extrapolation task, especially in their scores for the first presentation, in which no previous experience could be used. Laboratory rats and mice demonstrated a low level of extrapolation performance. This means that only a few laboratory animals were capable of solving the task, i.e., the percentage of correct solutions was equivalent to chance. The brain weight selection program resulted in two mice strains with a 20% (90-mg) difference in brain weight. Ability to solve the extrapolation task was present in low-brain weight mice in generations 7-11 but declined with further selection. Investigation of extrapolation ability in mice with different chromosomal anomalies demonstrated that animals with Robertsonian translocations Rb(8,17) 1lem and Rb(8,17) 6Sic were capable of solving this task in a statistically significant majority of cases, while mice with fusion of other chromosomes, as well as CBA normal karyotype mice, performed no better than expected by chance. Mice with two types of partial trisomies and animals homo- and heterozygous for translocations were also tested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8267558
Genetic Engineering of a Mouse
Jones, Dennis
Genetic engineering is the process of modifying an organismÆs genetic composition by adding foreign genes to produce desired traits or evaluate function. Dr. Jon W. Gordon and Sterling Professor Emeritus at Yale Dr. Frank H. Ruddle were pioneers in mammalian gene transfer research. Their research resulted in production of the first transgenic animals, which contained foreign DNA that was passed on to offspring. Transgenic mice have revolutionized biology, medicine, and biotechnology in the 21st century. In brief, this review revisits their creation of transgenic mice and discusses a few evolving applications of their transgenic technology used in biomedical research. PMID:21698043
Genetically Engineered Cyanobacteria
NASA Technical Reports Server (NTRS)
Zhou, Ruanbao (Inventor); Gibbons, William (Inventor)
The disclosed embodiments provide cyanobacteria spp. that have been genetically engineered to have increased production of carbon-based products of interest. These genetically engineered hosts efficiently convert carbon dioxide and light into carbon-based products of interest such as long chained hydrocarbons. Several constructs containing polynucleotides encoding enzymes active in the metabolic pathways of cyanobacteria are disclosed. In many instances, the cyanobacteria strains have been further genetically modified to optimize production of the carbon-based products of interest. The optimization includes both up-regulation and down-regulation of particular genes.
Progress and prospects: genetic engineering in xenotransplantation
S Le Bas-Bernardet; I Anegon; G Blancho
In this review, we summarize the work published over the last 2 years using genetic modifications of animals in the field of xenotransplantation. Genetic engineering of the donor has become a powerful tool in xenotransplantation, both for the inactivation of one particular porcine gene and for the addition of human genes with the goal of overcoming xenogeneic barriers. We summarize
Genetic Engineering in Floriculture
Yoshikazu Tanaka; Ryutaro Aida
\\u000a Numerous attractive floricultural crops have been developed by extensive hybridization and mutational breeding which suffer\\u000a from genetic constraint intrinsic to each plant species. Breeding by utilizing genetic engineering has liberated such constraint\\u000a and any genes from any organisms can be used to make novel floricultural crops. Novel violet\\/blue colored carnation and rose\\u000a have been developed by expressing flavonoid biosynthetic genes
Genetically Engineered Pig Models for Human Diseases
Prather, Randall S.; Lorson, Monique; Ross, Jason W.; Whyte, Jeffrey J.; Walters, Eric
Although pigs are used widely as models of human disease, their utility as models has been enhanced by genetic engineering. Initially, transgenes were added randomly to the genome, but with the application of homologous recombination, zinc finger nucleases, and transcription activator-like effector nuclease (TALEN) technologies, now most any genetic change that can be envisioned can be completed. To date these genetic modifications have resulted in animals that have the potential to provide new insights into human diseases for which a good animal model did not exist previously. These new animal models should provide the preclinical data for treatments that are developed for diseases such as Alzheimer's disease, cystic fibrosis, retinitis pigmentosa, spinal muscular atrophy, diabetes, and organ failure. These new models will help to uncover aspects and treatments of these diseases that were otherwise unattainable. The focus of this review is to describe genetically engineered pigs that have resulted in models of human diseases. PMID:25387017
Integration of Multi-objective and Interactive Genetic Algorithms and its Application to Animation Design
Coello, Carlos A. Coello
Integration of Multi-objective and Interactive Genetic Algorithms and its Application to Animation and Systems Science, Interdisciplinary Graduate School of Science and Engineering, Tokyo Institute human like motions in animation by computer graphics is a difficult task. Currently, motions
Introduction to Genetic Engineering and Its Applications
NSDL National Science Digital Library
National Science Foundation GK-12 and Research Experience for Teachers (RET) Programs,
Students learn how engineers apply their understanding of DNA to manipulate specific genes to produce desired traits, and how engineers have used this practice to address current problems facing humanity. They learn what genetic engineering means and examples of its applications, as well as moral and ethical problems related to its implementation. Students fill out a flow chart to list the methods to modify genes to create GMOs and example applications of bacteria, plant and animal GMOs.
Genetically Modified Animals and Pharmacological Research
Dominic J. Wells
\\u000a This chapter reviews the use of genetically modified animals and the increasingly detailed knowledge of the genomes of the\\u000a domestic species. The different approaches to genetic modification are outlined as are the advantages and disadvantages of\\u000a the techniques in different species. Genetically modified mice have been fundamental in understanding gene function and in\\u000a generating affordable models of human disease although
Genetic Approaches to Modeling Anxiety in Animals
Laura H. Jacobson; John F. Cryan
\\u000a Anxiety disorders are a growing health problem world-wide. However, the causative factors, etiology, and underlying mechanisms\\u000a of anxiety disorders, as for most psychiatric disorders, remain relatively poorly understood. The current status of clinical\\u000a research indicates that anxiety traits and anxiety disorder in man have a genetic component, and therefore genetic modeling\\u000a in animals is a logical approach to gain a
University Students' Knowledge and Attitude about Genetic Engineering
Nilay Keskin Samanci; Orńun Bozkurt
Genetic engineering and biotechnology made possible of gene transfer without discriminating microorganism, plant, animal or human. However, although these scientific techniques have benefits, they cause arguments because of their ethical and social impacts. The arguments about ethical ad social impacts of biotechnology made clear that not only getting basic knowledge about biotechnology and genetic engineering, also ethical and social issues
Animal Genetic Resource Trade Flows: Economic Assessment
Technology Transfer Automated Retrieval System (TEKTRAN)
Throughout human history, livestock producers have relied on a vibrant international exchange of genetic resources to achieve improvements in the quality and productivity of their animals. In recent years, however, some observers have argued that changes in the legal, technological, and economic env...
77 FR 41350 - Monsanto Co.; Determination of Nonregulated Status of Soybean Genetically Engineered To Produce...
Federal Register 2010, 2011, 2012, 2013, 2014
...Soybean Genetically Engineered To Produce Stearidonic Acid AGENCY: Animal and Plant Health Inspection Service...been genetically engineered to produce stearidonic acid, an omega-3 fatty acid not found in conventional soybean, is no longer...
Genetic engineering and the patent office
Jeffrey G. Sheldon; Denton L. Anderson
Higher life forms created through genetic engineering are now recognized as potentially patentable. On 7 April 1987, the US Patent and Trademark Office announced that it now considers non-naturally occurring non-human multi-cellular living organisms, including animals, to be patentable subject matter. The response to this announcement has been an emotion controversy centering on the patent office. The announcement has become
ASAS Centennial Paper: Future needs in animal breeding and genetics
R. D. Green
Abstract The past century has seen animal breeding and genetics evolve and expand from definition and validation of basic population genetics theory to development of selection index theory to today's relatively sophisticated genetic prediction systems enabling industry genetic improvement. The end of the first century of the American Society of Animal Science also coincides with the rapid movement,of the field
Animal Models for Adipose Tissue Engineering
Uthamanthil, Rajesh; Beahm, Elisabeth; Frye, Cindy
Abstract There is a critical need for adequate reconstruction of soft tissue defects resulting from tumor resection, trauma, and congenital abnormalities. To be sure, adipose tissue engineering strategies offer promising solutions. However, before clinical translation can occur, efficacy must be proven in animal studies. The aim of this review is to provide an overview of animal models currently employed for adipose tissue engineering. PMID:18544014
Genetic engineering of Geobacillus spp.
Kananavi?i?t?, R?ta; ?itavi?ius, Donaldas
Members of the genus Geobacillus are thermophiles that are of great biotechnological importance, since they are sources of many thermostable enzymes. Because of their metabolic versatility, geobacilli can be used as whole-cell catalysts in processes such as bioconversion and bioremediation. The effective employment of Geobacillus spp. requires the development of reliable methods for genetic engineering of these bacteria. Currently, genetic manipulation tools and protocols are under rapid development. However, there are several convenient cloning vectors, some of which replicate autonomously, while others are suitable for the genetic modification of chromosomal genes. Gene expression systems are also intensively studied. Combining these tools together with proper techniques for DNA transfer, some Geobacillus strains were shown to be valuable producers of recombinant proteins and industrially important biochemicals, such as ethanol or isobutanol. This review encompasses the progress made in the genetic engineering of Geobacillus spp. and surveys the vectors and transformation methods that are available for this genus. PMID:25659824
"Genetically Engineered" Nanoelectronics
NASA Technical Reports Server (NTRS)
Klimeck, Gerhard; Salazar-Lazaro, Carlos H.; Stoica, Adrian; Cwik, Thomas
The quantum mechanical functionality of nanoelectronic devices such as resonant tunneling diodes (RTDs), quantum well infrared-photodetectors (QWIPs), quantum well lasers, and heterostructure field effect transistors (HFETs) is enabled by material variations on an atomic scale. The design and optimization of such devices requires a fundamental understanding of electron transport in such dimensions. The Nanoelectronic Modeling Tool (NEMO) is a general-purpose quantum device design and analysis tool based on a fundamental non-equilibrium electron transport theory. NEW was combined with a parallelized genetic algorithm package (PGAPACK) to evolve structural and material parameters to match a desired set of experimental data. A numerical experiment that evolves structural variations such as layer widths and doping concentrations is performed to analyze an experimental current voltage characteristic. The genetic algorithm is found to drive the NEMO simulation parameters close to the experimentally prescribed layer thicknesses and doping profiles. With such a quantitative agreement between theory and experiment design synthesis can be performed.
Organic agriculture versus genetic engineering
H. Verhoog
The objections of organic agriculture against genetic engineering as presented in the 2002 Position Statement of the International Federation of Organic Agriculture Movements (IFOAM) are analysed. The objections can be grouped into three categories: risks to human health and the environment, socio-ethical objections, and incompatibility with the principles of sustainable agriculture. As to threats to human health and the environment
[Genetic behavioral aspects of agricultural animals].
Buchenauer, D
This paper reviews some experimental methods used in the study of genetic fixed behaviour, some examples of investigations in farm animals as well as some results of the author's studies on the emotionality in pigs are given. Many investigations have been made on the estimation of heritability of behaviour patterns in cattle, pigs, poultry and dogs. Behaviour patterns such as temperament, aggressiveness, and nervousness showed relatively high h2-values; other behaviours like trainability, emotionality, and pre-laying showed relatively low values. Selection experiments were carried out in poultry and dogs. The results showed that after a few generations remarkable differences in behaviour patterns between the new lines and the original populations were obvious. Besides the selected traits, modifications occurred in other behaviour patterns and in physiological responses. The emotionality in pigs was investigated with the open field test in 3 genetic groups: German Landrace (DL), Duroc (DU), and cross-breeding between these breeds (DU x DL). Increasing emotionality was displayed by increasing ambulatory activity and vocalization. The lowest activity was observed in the pure breeds, DU showed less signs of emotionality than DL, whereas DU x DL showed the highest of emotionality. PMID:2387229
Molecular Genetics of Ubiquinone Biosynthesis in Animals
Wang, Ying; Hekimi, Siegfried
Ubiquinone (UQ), also known as coenzyme Q (CoQ), is a redox-active lipid present in all cellular membranes where it functions in a variety of cellular processes. The best known functions of UQ are to act as a mobile electron carrier in the mitochondrial respiratory chain and to serve as a lipid soluble antioxidant in cellular membranes. All eukaryotic cells synthesize their own UQ. Most of the current knowledge on the UQ biosynthetic pathway was obtained by studying Escherichia coli and S. cerevisiae UQ-deficient mutants. The orthologues of all the genes known from yeast studies to be involved in UQ biosynthesis have subsequently been found in higher organisms. Animal mutants with different genetic defects in UQ biosynthesis display very different phenotypes, despite the fact that in all these mutants the same biosynthetic pathway is affected. This review summarizes the present knowledge of the eukaryotic biosynthesis of UQ, with focus on the biosynthetic genes identified in animals, including C. elegans, rodents and humans. Moreover, we review the phenotypes of mutants in these genes and discuss the functional consequences of UQ deficiency in general. PMID:23190198
Advances in genetic engineering of marine algae.
Qin, Song; Lin, Hanzhi; Jiang, Peng
Algae are a component of bait sources for animal aquaculture, and they produce abundant valuable compounds for the chemical industry and human health. With today's fast growing demand for algae biofuels and the profitable market for cosmetics and pharmaceuticals made from algal natural products, the genetic engineering of marine algae has been attracting increasing attention as a crucial systemic technology to address the challenge of the biomass feedstock supply for sustainable industrial applications and to modify the metabolic pathway for the more efficient production of high-value products. Nevertheless, to date, only a few marine algae species can be genetically manipulated. In this article, an updated account of the research progress in marine algal genomics is presented along with methods for transformation. In addition, vector construction and gene selection strategies are reviewed. Meanwhile, a review on the progress of bioreactor technologies for marine algae culture is also revisited. PMID:22634258
Genetic effects of harvest on wild animal populations
Genetic effects of harvest on wild animal populations Fred W. Allendorf1,2,3 , Phillip R. England4~o, Portugal 6 Division of Population Genetics, Department of Zoology, University of Stockholm, S-10691 the world and is often intense. Harvest has the potential to cause three types of genetic change: alteration
Animated Drawings Rendered by Genetic Programming Perry Barile
Ciesielski, Vic
Animated Drawings Rendered by Genetic Programming Perry Barile School of Computer Science describe an approach to generating animations of draw- ings that start as a random collection of strokes of an animation. In this paper we describe an approach to generating draw- ings of a kind that have not been done
Genetic Programming Evolution of Controllers for 3D Character Animation
Fernandez, Thomas
) by the animation software. The motion resulting from interpolating the key frames is purely kinematicGenetic Programming Evolution of Controllers for 3ŁD Character Animation Larry Gritz James K. Hahn Pixar Animation Studios 1001 W. Cutting Blvd. Richmond, CA 94804 email@example.com The George Washington
Animal Models for Bone Tissue Engineering Purposes
Vķronique Viateau; Delphine Logeart-Avramoglou; GeneviĶve Guillemin; Hervķ Petite
To assess the efficacy of engineered tissues, it is necessary to have (1) appropriate large animal models that mimic the clinical\\u000a setting and (2) relevant methods of monitoring the biofuntionality of these tissues. However, developing these tissue constructs\\u000a is a step-by-step process in which numerous variables such as scaffold design, source of stem cells and mode of growth factor\\u000a application
Enhanced Genetic Tools for Engineering Multigene Traits into Green Algae
Rasala, Beth A.; Chao, Syh-Shiuan; Pier, Matthew; Barrera, Daniel J.; Mayfield, Stephen P.
Transgenic microalgae have the potential to impact many diverse biotechnological industries including energy, human and animal nutrition, pharmaceuticals, health and beauty, and specialty chemicals. However, major obstacles to sophisticated genetic and metabolic engineering in algae have been the lack of well-characterized transformation vectors to direct engineered gene products to specific subcellular locations, and the inability to robustly express multiple nuclear-encoded transgenes within a single cell. Here we validate a set of genetic tools that enable protein targeting to distinct subcellular locations, and present two complementary methods for multigene engineering in the eukaryotic green microalga Chlamydomonas reinhardtii. The tools described here will enable advanced metabolic and genetic engineering to promote microalgae biotechnology and product commercialization. PMID:24710110
Reproductive biotechnologies and management of animal genetic resources
Technology Transfer Automated Retrieval System (TEKTRAN)
Global awareness has increased efforts to conserve animal genetic resources (AnGR). Ex-situ conservation and management of AnGR is exclusively dependent upon an array of reproductive and genetic biotechnologies. These technologies range from well established protocols, e.g., cryopreservation of sper...
Considerations for the assessment of the safety of genetically modified animals used for human food or animal feed
Gijs A Kleter; Harry A Kuiper
Genetically modified food and feed crops have entered the Western market, and genetically modified animals may follow in the near future. The issues that are commonly addressed in the assessment of the safety of genetically modified crops are discussed, as well as the analogous issues that may arise for genetically modified animals. For safety assessment, the degree of substantial equivalence
Advanced Animation Engine for User-Interface Robots
Albert J. N. Van Breemen; Yan Xue
This paper describes an improved animation system for user-interface robots. The animation system is based on the animation engine presented by van Breemen, A.J.N. (2004), and uses animation channels for playing and blending multiple animations concurrently. The presented improvement is twofold. First, this paper describes an extension to the computational structure of an animation channel. A fading mechanism is added
Fast Physical Simulation Engines for Creating Dynamic Character Animations
Katsu YAMANE; Masaya HIRASHIMA; Yoshihiko NAKAMURA
This paper presents our computational engines for physical simulation of human\\/animal characters, which are useful for synthesizing dynamic character animations. In particular, we focus on their extension to inverse dynamics calculation of musculoskeletal human model. Such model would be essential to future animation contents that require detailed and realistic characters. Keyword Character Animation, Musculoskeletal Model, Inverse Dynamics, Dynamics Simulation
Precision genetic engineering in large mammals.
Garrels, Wiebke; Ivics, Zoltan; Kues, Wilfried A
Precision genetic engineering based on stable chromosomal insertion of exogenous DNA in the genomes of large mammals is immensely important for the development of improved biomedical models, pharmaceutical research and an accelerated breeding progress. Precision genetic engineering requires (i) a known locus of genomic integration, (ii) a defined status of foreign DNA, (iii) that transgene expression is unaffected by neighbouring chromosomal sequences, (iv) endogenous genes are not mutated and (v) no unwanted DNA sequences are present. Recently, advanced molecular techniques exploiting exogenous enzymes have opened the possibilities for more sophisticated genetic engineering. Here, we critically review current developments of enzyme-catalysed approaches for targeted transgenesis in large mammals. PMID:22521716
Pertussis toxins, other antigens become likely targets for genetic engineering
Marwick, C.
Genetically engineered pertussis vaccines have yet to be fully tested clinically. But early human, animal, and in vitro studies indicate effectiveness in reducing toxic effects due to Bordetella pertussis. The licensed pertussis vaccines consists of inactivated whole cells of the organism. Although highly effective, they have been associated with neurologic complications. While the evidence continues to mount that these complications are extremely rare, if they occur at all, it has affected the public's acceptance of pertussis immunization.
Genetically modified ingredients in animal nutrition: Their safety and future
A. Chesson; H. J. Flint
SUMMARY -The immense potential of genetic manipulation techniques is now being realized with a dramatic increase in the agricultural and industrial use of modified plants and microorganisms. Many animal feeds now include material from crop plants that have been modified for characteristics such as disease or pest resistance that are unlikely to affect their nutritional value. In addition crop plants
From genetical genomics to systems genetics: potential applications in quantitative genomics and animal breeding
Haja N. Kadarmideen; Peter von Rohr; Luc L. G. Janss
This article reviews methods of integration of transcriptomics (and equally proteomics and metabolomics), genetics, and genomics\\u000a in the form of systems genetics into existing genome analyses and their potential use in animal breeding and quantitative\\u000a genomic modeling of complex traits. Genetical genomics or the expression quantitative trait loci (eQTL) mapping method and\\u000a key findings in this research are reviewed. Various
Genetically Engineered Immunotherapy for Advanced Cancer
In this trial, doctors will collect T lymphocytes from the blood of patients with advanced mesothelin-expressing cancer and genetically engineer these cells to recognize mesothelin. The gene-engineered cells will then be multiplied and infused into the patients to fight their cancer.
DNA marker technology: a revolution in animal genetics.
Dodgson, J B; Cheng, H H; Okimoto, R
The development of DNA-based markers has had a revolutionary impact on gene mapping and, more generally, on all of animal and plant genetics. With DNA-based markers, it is theoretically possible to exploit the entire diversity in DNA sequence that exists in any cross. For this reason, high resolution genetic maps are being developed at an unprecedented speed. The most commonly used DNA-based markers include those based on a cloned and (usually) sequenced DNA fragment and other, more random, assays for genetic polymorphism that can be grouped under the heading of fingerprint markers. The advantages and disadvantages of the various marker types are discussed, along with their application to the reference chicken genetic linkage maps and to the search for quantitative trait loci (QTL). The prospects for the use of DNA-based markers in marker-assisted selection are considered, along with likely future trends in poultry gene mapping. Further development of both physical and linkage genome maps of the chicken will allow animal scientists to more efficiently detect and characterize QTL and will provide them access to the wealth of genetic information that is being generated about the human genome and the genomes of model species, such as the mouse and Drosophila. PMID:9251136
Potency of Animal Models in KANSEI Engineering
NASA Astrophysics Data System (ADS)
Ozaki, Shigeru; Hisano, Setsuji; Iwamoto, Yoshiki
Various species of animals have been used as animal models for neuroscience and provided critical information about the brain functions. Although it seems difficult to elucidate a highly advanced function of the human brain, animal models have potency to clarify the fundamental mechanisms of emotion, decision-making and social behavior. In this review, we will pick up common animal models and point to both the merits and demerits caused by the characteristics. We will also mention that wide-ranging approaches from animal models are advantageous to understand KANSEI as well as mind in humans.
Genetically engineered nanocarriers for drug delivery
Shi, Pu; Gustafson, Joshua A; MacKay, J Andrew
Cytotoxicity, low water solubility, rapid clearance from circulation, and off-target side-effects are common drawbacks of conventional small-molecule drugs. To overcome these shortcomings, many multifunctional nanocarriers have been proposed to enhance drug delivery. In concept, multifunctional nanoparticles might carry multiple agents, control release rate, biodegrade, and utilize target-mediated drug delivery; however, the design of these particles presents many challenges at the stage of pharmaceutical development. An emerging solution to improve control over these particles is to turn to genetic engineering. Genetically engineered nanocarriers are precisely controlled in size and structure and can provide specific control over sites for chemical attachment of drugs. Genetically engineered drug carriers that assemble nanostructures including nanoparticles and nanofibers can be polymeric or non-polymeric. This review summarizes the recent development of applications in drug and gene delivery utilizing nanostructures of polymeric genetically engineered drug carriers such as elastin-like polypeptides, silk-like polypeptides, and silk-elastin-like protein polymers, and non-polymeric genetically engineered drug carriers such as vault proteins and viral proteins. PMID:24741309
Genetic Engineering Strategies for Enhanced Biodiesel Production.
Hegde, Krishnamoorthy; Chandra, Niharika; Sarma, Saurabh Jyoti; Brar, Satinder Kaur; Veeranki, Venkata Dasu
The focus on biodiesel research has shown a tremendous growth over the last few years. Several microbial and plant sources are being explored for the sustainable biodiesel production to replace the petroleum diesel. Conventional methods of biodiesel production have several limitations related to yield and quality, which led to development of new engineering strategies to improve the biodiesel production in plants, and microorganisms. Substantial progress in utilizing algae, yeast, and Escherichia coli for the renewable production of biodiesel feedstock via genetic engineering of fatty acid metabolic pathways has been reported in the past few years. However, in most of the cases, the successful commercialization of such engineering strategies for sustainable biodiesel production is yet to be seen. This paper systematically presents the drawbacks in the conventional methods for biodiesel production and an exhaustive review on the present status of research in genetic engineering strategies for production of biodiesel in plants, and microorganisms. Further, we summarize the technical challenges need to be tackled to make genetic engineering technology economically sustainable. Finally, the need and prospects of genetic engineering technology for the sustainable biodiesel production and the recommendations for the future research are discussed. PMID:25902752
The value of animal models in predicting genetic susceptibility to complex diseases such as rheumatoid arthritis
Emma Ahlqvist; Malin Hultqvist; Rikard Holmdahl
For a long time, genetic studies of complex diseases were most successfully conducted in animal models. However, the field of genetics is now rapidly evolving, and human genetics has also started to produce strong candidate genes for complex diseases. This raises the question of how to continue gene-finding attempts in animals and how to use animal models to enhance our
Molecular scissors and their application in genetically modified farm animals.
Petersen, Bjoern; Niemann, Heiner
Molecular scissors (MS), incl. Zinc Finger Nucleases (ZFN), Transcription-activator like endoncleases (TALENS) and meganucleases possess long recognition sites and are thus capable of cutting DNA in a very specific manner. These molecular scissors mediate targeted genetic alterations by enhancing the DNA mutation rate via induction of double-strand breaks at a predetermined genomic site. Compared to conventional homologous recombination based gene targeting, MS can increase the targeting rate 10,000-fold, and gene disruption via mutagenic DNA repair is stimulated at a similar frequency. The successful application of different MS has been shown in different organisms, including insects, amphibians, plants, nematodes, and mammals, including humans. Recently, another novel class of molecular scissors was described that uses RNAs to target a specific genomic site. The CRISPR/Cas9 system is capable of targeting even multiple genomic sites in one shot and thus could be superior to ZFNs or TALEN, especially by its easy design. MS can be successfully employed for improving the understanding of complex physiological systems, producing transgenic animals, incl. creating large animal models for human diseases, creating specific cell lines, and plants, and even for treating human genetic diseases. This review provides an update on molecular scissors, their underlying mechanism and focuses on new opportunities for generating genetically modified farm animals. PMID:25603988
Recent developments in the genetic engineering of barley
Mannonen, L.; Kauppinen, V.; Enari, T.M. (VTT Biotechnology and Food Research, Espoo (Finland))
Cereals are the most important group of plants for human nutrition and animal feed. Partially due to the commercial value of crop plants, there has been an ever-increasing interest in using modern biotechnological methods for the improvement of the characteristics of cereals during the past decade. The rapid progress in molecular biology, plant cell culture techniques, and gene transfer technology has resulted in successful transformations of all the major cereals--maize, rice, wheat, and barley. This brings the biotechnological methods closer to the routine also in barley breeding. In this article, the current status of barley genetic engineering, including the patent situation, is reviewed. The needs aims, and possible applications of genetic engineering in barley breeding are discussed. 179 refs.
Genetic engineering of Mycobacterium tuberculosis: a review.
Lamrabet, Otmane; Drancourt, Michel
Genetic engineering has been used for decades to mutate and delete genes in the Mycobacterium tuberculosis genome with the translational goal of producing attenuated mutants with conserved susceptibility to antituberculous antibiotics. The development of plasmids and mycobacteriophages that can transfer DNA into the M. tuberculosis chromosome has effectively overcome M. tuberculosis slow growth rate and the capsule and mycolic acid wall, which limit DNA uptake. The use of genetic engineering techniques has shed light on many aspects of pathogenesis mechanisms, including cellular growth, mycolic acid biosynthesis, metabolism, drug resistance and virulence. Moreover, such research gave clues to the development of new vaccines or new drugs for routine clinical practice. The use of genetic engineering tools is mainly based on the underlying concept that altering or reducing the M. tuberculosis genome could decrease its virulence. A contrario, recent post-genomic analyses indicated that reduced bacterial genomes are often associated with increased bacterial virulence and that M. tuberculosis acquired genes by lateral genetic exchange during its evolution. Therefore, ancestors utilizing genetic engineering to add genes to the M. tuberculosis genome may lead to new vaccines and the availability of M. tuberculosis isolates with increased susceptibility to antituberculous antibiotics. PMID:22789498
Of mice and microflora: considerations for genetically engineered mice.
Treuting, P M; Clifford, C B; Sellers, R S; Brayton, C F
The phenotype of genetically engineered mice is a combination of both genetic and environmental factors that include the microflora of the mouse. The impact a particular microbe has on a mouse reflects the host-microbe interaction within the context of the mouse genotype and environment. Although often considered a confounding variable, many host-microbe interactions have resulted in the generation of novel model systems and characterization of new microbial agents. Microbes associated with overt disease in mice have been the historical focus of the laboratory animal medical and pathology community and literature. The advent of genetic engineering and the complex of mouse models have revealed previously unknown or disregarded agents that now oblige the attention of the biomedical research community. The purpose of this article is to describe and illustrate how phenotypes can be affected by microflora by focusing on the infectious diseases present in genetically engineered mouse (GEM) colonies of our collective institutions and by reviewing important agents that are rarely seen in most research facilities today. The goal is to introduce the concept of the role of microflora on phenotypes and in translational research using GEM models. PMID:22173977
Genetic Engineering of Allergens: Future Therapeutic Products
Fßtima Ferreira; Michael Wallner; Heimo Breiteneder; Arnulf Hartl; Josef Thalhamer; Christof Ebner
Genetic engineering of allergens for specific immunotherapy should aim at the production of modified molecules with reduced IgE-binding epitopes (hypoallergens), while preserving structural motifs necessary for T cell recognition (T cell epitopes) and for induction of IgG antibodies reactive with the natural allergen (blocking antibodies). Common approaches for engineering of hypoallergens usually require knowledge of T and B cell epitopes
Animal Models for Vascular Tissue-Engineering
Swartz, Daniel D.; Andreadis, Stelios T.
Due to rise in cardiovascular disease throughout the world, there is increasing demand for small diameter blood vessels as replacement grafts. The present review focuses on the animal models that have been used to test small-diameter TEVs with emphasis on the attributes of each model. Small animal models are used to test short-term patency and address mechanistic hypotheses; and large, pre-clinical animal models are employed to test long-term patency, remodeling and function in an environment mimicking human physiology. We also discuss recent clinical trials that employed laboratory fabricated TEVs and showed very promising results. Ultimately, animal models provide a testing platform for optimizing vascular grafts before clinical use in patients without suitable autologous vessels. PMID:23769861
Genetically Engineered Biologically Based Hemostatic Bioassay
Lilong Tang; David J. Christini; Jay M. Edelberg
Real-time direct measures of hemostatic parameters in vivo are required for optimizing the dynamic delivery of coagulation modifying pharmacotherapies. Typical sensors of physiologic functions in vivo, however, have only a restricted array of sensory inputs, and thus limited capacity to monitor thrombotic and hemostatic activity. To overcome this limitation we have developed a genetically engineered excitable cell line that can
Integrated Pest Management and Genetically Engineered Plants
G. Sch³tte
Summary 1. EU directives lay down, that genetically engineered (GE) organisms should neither cause direct nor indirect negative (acute or long-term) effects and pesticide use should be founded on the principles of integrated pest management (IPM). IPM is an important international political goal. 2. Results of field tests and studies on integrated farming and on the agricultural practice in GE
Specific genetic modifications of domestic animals by gene targeting and animal cloning.
Wang, Bin; Zhou, Jiangfeng
The technology of gene targeting through homologous recombination has been extremely useful for elucidating gene functions in mice. The application of this technology was thought impossible in the large livestock species until the successful creation of the first mammalian clone "Dolly" the sheep. The combination of the technologies for gene targeting of somatic cells with those of animal cloning made it possible to introduce specific genetic mutations into domestic animals. In this review, the principles of gene targeting in somatic cells and the challenges of nuclear transfer using gene-targeted cells are discussed. The relevance of gene targeting in domestic animals for applications in bio-medicine and agriculture are also examined. PMID:14614774
Specific genetic modifications of domestic animals by gene targeting and animal cloning
Wang, Bin; Zhou, Jiangfeng
The technology of gene targeting through homologous recombination has been extremely useful for elucidating gene functions in mice. The application of this technology was thought impossible in the large livestock species until the successful creation of the first mammalian clone "Dolly" the sheep. The combination of the technologies for gene targeting of somatic cells with those of animal cloning made it possible to introduce specific genetic mutations into domestic animals. In this review, the principles of gene targeting in somatic cells and the challenges of nuclear transfer using gene-targeted cells are discussed. The relevance of gene targeting in domestic animals for applications in bio-medicine and agriculture are also examined. PMID:14614774
Genetic recombination between human and animal parasites creates novel strains of human pathogen.
Gibson, Wendy; Peacock, Lori; Ferris, Vanessa; Fischer, Katrin; Livingstone, Jennifer; Thomas, James; Bailey, Mick
Genetic recombination between pathogens derived from humans and livestock has the potential to create novel pathogen strains, highlighted by the influenza pandemic H1N1/09, which was derived from a re-assortment of swine, avian and human influenza A viruses. Here we investigated whether genetic recombination between subspecies of the protozoan parasite, Trypanosoma brucei, from humans and animals can generate new strains of human pathogen, T. b. rhodesiense (Tbr) responsible for sleeping sickness (Human African Trypanosomiasis, HAT) in East Africa. The trait of human infectivity in Tbr is conferred by a single gene, SRA, which is potentially transferable to the animal pathogen Tbb by sexual reproduction. We tracked the inheritance of SRA in crosses of Tbr and Tbb set up by co-transmitting genetically-engineered fluorescent parental trypanosome lines through tsetse flies. SRA was readily transferred into new genetic backgrounds by sexual reproduction between Tbr and Tbb, thus creating new strains of the human pathogen, Tbr. There was no evidence of diminished growth or transmissibility of hybrid trypanosomes carrying SRA. Although expression of SRA is critical to survival of Tbr in the human host, we show that the gene exists as a single copy in a representative collection of Tbr strains. SRA was found on one homologue of chromosome IV in the majority of Tbr isolates examined, but some Ugandan Tbr had SRA on both homologues. The mobility of SRA by genetic recombination readily explains the observed genetic variability of Tbr in East Africa. We conclude that new strains of the human pathogen Tbr are being generated continuously by recombination with the much larger pool of animal-infective trypanosomes. Such novel recombinants present a risk for future outbreaks of HAT. PMID:25816228
Genetic Recombination between Human and Animal Parasites Creates Novel Strains of Human Pathogen
Gibson, Wendy; Peacock, Lori; Ferris, Vanessa; Fischer, Katrin; Livingstone, Jennifer; Thomas, James; Bailey, Mick
Genetic recombination between pathogens derived from humans and livestock has the potential to create novel pathogen strains, highlighted by the influenza pandemic H1N1/09, which was derived from a re-assortment of swine, avian and human influenza A viruses. Here we investigated whether genetic recombination between subspecies of the protozoan parasite, Trypanosoma brucei, from humans and animals can generate new strains of human pathogen, T. b. rhodesiense (Tbr) responsible for sleeping sickness (Human African Trypanosomiasis, HAT) in East Africa. The trait of human infectivity in Tbr is conferred by a single gene, SRA, which is potentially transferable to the animal pathogen Tbb by sexual reproduction. We tracked the inheritance of SRA in crosses of Tbr and Tbb set up by co-transmitting genetically-engineered fluorescent parental trypanosome lines through tsetse flies. SRA was readily transferred into new genetic backgrounds by sexual reproduction between Tbr and Tbb, thus creating new strains of the human pathogen, Tbr. There was no evidence of diminished growth or transmissibility of hybrid trypanosomes carrying SRA. Although expression of SRA is critical to survival of Tbr in the human host, we show that the gene exists as a single copy in a representative collection of Tbr strains. SRA was found on one homologue of chromosome IV in the majority of Tbr isolates examined, but some Ugandan Tbr had SRA on both homologues. The mobility of SRA by genetic recombination readily explains the observed genetic variability of Tbr in East Africa. We conclude that new strains of the human pathogen Tbr are being generated continuously by recombination with the much larger pool of animal-infective trypanosomes. Such novel recombinants present a risk for future outbreaks of HAT. PMID:25816228
Ammonia Emissions and Animal Agriculture Susan W. Gay, Extension Engineer, Biological Systems Engineering, Virginia Tech
Liskiewicz, Maciej
Ammonia Emissions and Animal Agriculture Susan W. Gay, Extension Engineer, Biological Systems Engineering, Virginia Tech Katharine F. Knowlton, Assistant Professor, Dairy Science, Virginia Tech during the past two decades, air quality issues have become an increasing concern. Odors have been
Genetic control of programmed cell death during animal development
Conradt, Barbara
The elimination by programmed cell death of æunwantedÆ cells is a common feature of animal development. Genetic studies in the nematode Caenorhabditis elegans, the fruit fly Drosophila melanogaster and the mouse have not only revealed the molecular machineries that cause the programmed demise of specific cells, but have also allowed us to get a glimpse of the types of pathways that regulate these machineries during development. Rather than giving a broad overview of programmed cell death during development, the current review will focus on recent advances in our understanding of the regulation of specific programmed cell death events during nematode, fly and mouse development. These studies have revealed that many of the regulatory pathways involved have additional important roles in development, which confirms that the programmed cell death fate is an integral aspect of animal development. PMID:19886811
Animal Models for the Evaluation of Tissue Engineering Constructs
Daniel A. W. Oortgiesen; Gert J. Meijer; Rob B. M. Vries; X. Frank Walboomers; John A. Jansen
\\u000a In the last decade, tissue engineering has attracted a considerable amount of attention in medical research. Obviously, tissue-engineered\\u000a constructs need to be tested for their safety and efficacy before they can be used in the daily clinic. At present, animal\\u000a models offer the best possibility to do so. Each medical specialty favors its own specific model to test tissue-engineered\\u000a constructs.
Attitudes Toward Genetic Engineering: The Dilemma of the Genetically Abnormal Child
Gay Young; Cherylon Robinson
Ethicists and scientists have grappled with issues of genetic engineering for years, yet the position of the lay public on this topic remains largely unexplored. This study examines the attitudes of potential consumers Ś i.e., women of childbearing age Ś toward seven medical procedures for genetic engineering. We define genetic engineering as the use of medical procedures to terminate the
Genetic algorithms applied to optics and engineering
NASA Astrophysics Data System (ADS)
Cuevas, Francisco; Gonzalez, Otoniel; Susuki, Yamily; Hernandez, Daniel; Rocha, Martha; Alcala, Noķ
In the last years, Soft computing techniques, such as Genetic Algorithms, Neural Networks and Fuzzy systems, have been applied in different science areas. In this work, two applications of Genetic Algorithms in engineering and optics are presented. The Genetic Algorithms are optimization, search and learning machine techniques, which work in a random way. To achieve the problem solution by using of Genetic Algorithms, an iterative process should be developed. First, the problem to solve is modelled in a mathematical way by establishing of a fitness or objective function. After, a random initial population of strings (chromosomes) codifying problem solutions is generated, which samples the search solution space of the fitness function. Then, offspring populations are generated from previous one by using genetic operators: selection, crossover and mutation. In the selection process, possible solutions are chosen depending on their fitness function value. Then, in the crossover procedure, string segments of pairs of solutions are exchanged to generate the next population. Finally, some parameters in the offspring population are changed by mutation with a low probability. Results of the application of Genetic Algorithms to solve fringe analysis and nesting in finite materials problems are presented.
Regulatory and Biosafety Issues in Relation to Transgenic Animals in Food and Agriculture, Feeds Containing Genetically Modified Organisms (GMO) and Veterinary Biologics
H. P. S. Kochhar; G. A. Gifford; S. Kahn
Development of an effective regulatory system for genetically engineered animals and their products has been the subject of increasing discussion among researchers, industry and policy developers, as well as the public. Since transgenesis and cloning are relatively new scientific techniques, transgenic animals are new organisms for which there is limited information. The issues associated with the regulation and biosafety of
Genetic engineering of microalgae for fuel production
Terri G. D³nahay; Eric E. Jarvis; Kathryn G. Zeiler; Paul G. Roessler; Lewis M. Brown
SummaryĀĀSignificant progress has been made toward the successful genetic engineering of microalgal species with high potential for\\u000a fuel production. Foreign DNA has been transferred into a green alga,Chlorella ellipsoidea, and has been successfully expressed in this heterologous system. In addition, electroporation has shown promise as a means\\u000a of introducing DNA into intact algal cells. We have analyzed the composition of
Designing and engineering evolutionary robust genetic circuits
Background One problem with engineered genetic circuits in synthetic microbes is their stability over evolutionary time in the absence of selective pressure. Since design of a selective environment for maintaining function of a circuit will be unique to every circuit, general design principles are needed for engineering evolutionary robust circuits that permit the long-term study or applied use of synthetic circuits. Results We first measured the stability of two BioBrick-assembled genetic circuits propagated in Escherichia coli over multiple generations and the mutations that caused their loss-of-function. The first circuit, T9002, loses function in less than 20 generations and the mutation that repeatedly causes its loss-of-function is a deletion between two homologous transcriptional terminators. To measure the effect between transcriptional terminator homology levels and evolutionary stability, we re-engineered six versions of T9002 with a different transcriptional terminator at the end of the circuit. When there is no homology between terminators, the evolutionary half-life of this circuit is significantly improved over 2-fold and is independent of the expression level. Removing homology between terminators and decreasing expression level 4-fold increases the evolutionary half-life over 17-fold. The second circuit, I7101, loses function in less than 50 generations due to a deletion between repeated operator sequences in the promoter. This circuit was re-engineered with different promoters from a promoter library and using a kanamycin resistance gene (kanR) within the circuit to put a selective pressure on the promoter. The evolutionary stability dynamics and loss-of-function mutations in all these circuits are described. We also found that on average, evolutionary half-life exponentially decreases with increasing expression levels. Conclusions A wide variety of loss-of-function mutations are observed in BioBrick-assembled genetic circuits including point mutations, small insertions and deletions, large deletions, and insertion sequence (IS) element insertions that often occur in the scar sequence between parts. Promoter mutations are selected for more than any other biological part. Genetic circuits can be re-engineered to be more evolutionary robust with a few simple design principles: high expression of genetic circuits comes with the cost of low evolutionary stability, avoid repeated sequences, and the use of inducible promoters increases stability. Inclusion of an antibiotic resistance gene within the circuit does not ensure evolutionary stability. PMID:21040586
Moral imagination in tissue engineering research on animal models
Anders Nordgren
Animal experimentation is an integrated part of tissue engineering research. This paper investigates what scientists conducting such experimentation should reasonably take into consideration from an ethical point of view. It is argued that scientists should use their moral imagination in making fundamental ethical choices, in reflecting on legal regulation, in taking public opinion seriously, and in balancing human benefit and
Greta: A Simple Facial Animation Engine Stefano Pasquariello1
Pelachaud, Catherine
the dynamics aspect of the human face. We have realized a Simple Facial Animation Engine (SFAE) where the 3D a pseudo-muscular approach to emulate the behaviour of face tissues and also includes particular features such as wrinkles and furrow to enhance its realism. In particular, the wrinkles have been implemented using bump
Genetic engineering of microorganisms for biodiesel production
Lin, Hui; Wang, Qun; Shen, Qi; Zhan, Jumei; Zhao, Yuhua
Biodiesel, as one type of renewable energy, is an ideal substitute for petroleum-based diesel fuel and is usually made from triacylglycerides by transesterification with alcohols. Biodiesel production based on microbial fermentation aiming to establish more efficient, less-cost and sustainable biodiesel production strategies is under current investigation by various start-up biotechnology companies and research centers. Genetic engineering plays a key role in the transformation of microbes into the desired cell factories with high efficiency of biodiesel production. Here, we present an overview of principal microorganisms used in the microbial biodiesel production and recent advances in metabolic engineering for the modification required. Overexpression or deletion of the related enzymes for de novo synthesis of biodiesel is highlighted with relevant examples. PMID:23222170
Natural and Genetically Engineered Proteins for Tissue Engineering
Gomes, SĒlvia; Leonor, Isabel B.; Mano, JoŃo F.; Reis, Rui L.
To overcome the limitations of traditionally used autografts, allografts and, to a lesser extent, synthetic materials, there is the need to develop a new generation of scaffolds with adequate mechanical and structural support, control of cell attachment, migration, proliferation and differentiation and with bio-resorbable features. This suite of properties would allow the body to heal itself at the same rate as implant degradation. Genetic engineering offers a route to this level of control of biomaterial systems. The possibility of expressing biological components in nature and to modify or bioengineer them further, offers a path towards multifunctional biomaterial systems. This includes opportunities to generate new protein sequences, new self-assembling peptides or fusions of different bioactive domains or protein motifs. New protein sequences with tunable properties can be generated that can be used as new biomaterials. In this review we address some of the most frequently used proteins for tissue engineering and biomedical applications and describe the techniques most commonly used to functionalize protein-based biomaterials by combining them with bioactive molecules to enhance biological performance. We also highlight the use of genetic engineering, for protein heterologous expression and the synthesis of new protein-based biopolymers, focusing the advantages of these functionalized biopolymers when compared with their counterparts extracted directly from nature and modified by techniques such as physical adsorption or chemical modification. PMID:22058578
Engineering control of airborne disease transmission in animal laboratories.
Kowalski, W J; Bahnfleth, W P; Carey, D D
We here present a review of the problem of controlling airborne disease transmission in animal research facilities, with emphasis on engineering design and air-treatment technologies. Dilution ventilation, pressurization control, source control, and air disinfection and removal systems are reviewed, and analytical studies on the effects of dilution ventilation, filtration, and ultraviolet germicidal irradiation are summarized. In addition, we discuss practical problems common to laboratory facilities and present a database of potential airborne pathogens and allergens that can be transmitted between humans and animals. We offer some conclusions regarding the design and selection of available technologies and components and provide cost estimates for various air-cleaning systems. PMID:12051655
Computing genetic evaluations through application of generalized least squares to an animal model
Paris-Sud XI, Universitķ de
Computing genetic evaluations through application of generalized least squares to an animal model G.F.S. HUDSON Department of Animal Sciences, University of Maryland, College Park, Maryland 20742, U.S.A. Summary The animal model for performance data is rewritten in the form of a fixed model with uncorrelated
Social Movement Organizations' Reactions to Genetic Engineering in Agriculture
Numerous social movement organizations are actively opposing genetic engineering in agriculture. This article looks at a coalition of movement groups opposing biotechnology and (b) the leading U.S. advocacy groups to determine the breadth of movement resistance. Movements resisting genetic engineering are acting consistently with their previous positions on issues, indicating a high degree of narrative fidelity between belief and action.
Lecturers and Academic Staff: Department of Biotechnology and Genetic Engineering
Lecturers and Academic Staff: Department of Biotechnology and Genetic Engineering Full-Time Total Technicians and Teaching Assistants The Departments of Biotechnology and Genetic Engineering have a number to provide the local and regional markets with highly qualified graduates in Mathematics, and biotechnology
Engineering plants for animal feed for improved nutritional value.
Williams, Peter E V
Feed formulation to meet nutritional requirements of livestock is becoming increasingly challenging. Regulations have banned the use of traditional high-quality protein supplements such as meat-and-bone meal, pollution from animal excreta of N and P is an issue and antibiotics are no longer available as insurance against the impact of enteric infection and feed anti-nutritional factors. The improved genetic potential of livestock is increasing daily requirement for energy and protein (essential amino acids). To benefit from the enhanced growth potential of livestock diets with high nutrient density are needed that can be formulated from crops without increased cost. Genetic modification of commodity crops used to manufacture animal feed in order to improve the density and quality of available nutrients is a potential solution to some of these problems. Furthermore, crops may be used as biofactories to produce molecules and products used in animal feed with considerable reductions in manufacturing fixed costs. Nevertheless, there are considerable not insurmountable challenges, such as the creation of sufficient economic value to deliver benefit to all members in the feed production chain, which is an essential element of identity preserving and delivering the technology to livestock producers. Individual output traits in the major commodity crops may not provide sufficient value to adequately compensate all the members of the feed production chain. Successful adoption of output traits may rely on inserting combinations of agronomic input traits with specific quality traits or increasing the value proposition by inserting combinations of output traits. PMID:14506877
Engineering in the biological substrate: information processing in genetic circuits
We review the rapidly evolving efforts to analyze, model, simulate, and engineer genetic and biochemical information processing systems within living cells. We begin by showing that the fundamental elements of information processing in electronic and genetic systems are strikingly similar, and follow this theme through a review of efforts to create synthetic genetic circuits. In particular, we describe and review
Genospirituality: genetic engineering for spiritual and religious enhancement.
Charlton, Bruce G
The most frequently discussed role for genetic engineering is in relation to medicine, and a second area which provokes discussion is the use of genetic engineering as an enhancement technology. But one neglected area is the potential use of genetic engineering to increase human spiritual and religious experience - or genospirituality. If technologies are devised which can conveniently and safely engineer genes causal of spiritual and religious behaviours, then people may become able to choose their degree of religiosity or spiritual sensitivity. For instance, it may become possible to increase the likelihood of direct religious experience - i.e. 'revelation': the subjective experience of communication from the deity. Or, people may be able to engineer 'animistic' thinking, a mode of cognition in which the significant features of the world - such as large animals, trees, distinctive landscape features - are regarded as sentient and intentional beings; so that the individual experiences a personal relationship with the world. Another potentially popular spiritual ability would probably be shamanism; in which states of altered consciousness (e.g. trances, delirium or dreams) are induced and the shaman may undergo the experience of transformations, 'soul journeys' and contact with a spirit realm. Ideally, shamanistic consciousness could be modulated such that trances were self-induced only when wanted and when it was safe and convenient; and then switched-off again completely when full alertness and concentration are necessary. It seems likely that there will be trade-offs for increased spirituality; such as people becoming less 'driven' to seek status and monetary rewards - as a result of being more spiritually fulfilled people might work less hard and take more leisure. On the other hand, it is also possible that highly moral, altruistic, peaceable and principled behaviours might become more prevalent; and the energy and joyousness of the best churches might spread and be strengthened. Overall, genospirituality would probably be used by people who were unable to have the kind of spiritual or religious experiences which they wanted (or perhaps even needed) in order to lead the kind of life to which they aspired. PMID:18782654
Agrobacterium: natureÆs genetic engineer
Nester, Eugene W.
Agrobacterium was identified as the agent causing the plant tumor, crown gall over 100 years ago. Since then, studies have resulted in many surprising observations. Armin Braun demonstrated that Agrobacterium infected cells had unusual nutritional properties, and that the bacterium was necessary to start the infection but not for continued tumor development. He developed the concept of a tumor inducing principle (TIP), the factor that actually caused the disease. Thirty years later the TIP was shown to be a piece of a tumor inducing (Ti) plasmid excised by an endonuclease. In the next 20 years, most of the key features of the disease were described. The single-strand DNA (T-DNA) with the endonuclease attached is transferred through a type IV secretion system into the host cell where it is likely coated and protected from nucleases by a bacterial secreted protein to form the T-complex. A nuclear localization signal in the endonuclease guides the transferred strand (T-strand), into the nucleus where it is integrated randomly into the host chromosome. Other secreted proteins likely aid in uncoating the T-complex. The T-DNA encodes enzymes of auxin, cytokinin, and opine synthesis, the latter a food source for Agrobacterium. The genes associated with T-strand formation and transfer (vir) map to the Ti plasmid and are only expressed when the bacteria are in close association with a plant. Plant signals are recognized by a two-component regulatory system which activates vir genes. Chromosomal genes with pleiotropic functions also play important roles in plant transformation. The data now explain BraunÆs old observations and also explain why Agrobacterium is natureÆs genetic engineer. Any DNA inserted between the border sequences which define the T-DNA will be transferred and integrated into host cells. Thus, Agrobacterium has become the major vector in plant genetic engineering. PMID:25610442
Maintenance of quantitative genetic variation in animal breeding programmes
William G Hill
Factors influencing the maintenance of genetic variation in quantitative traits in populations undergoing artificial selection are reviewed. Formulae are given for simple cases, in particular for the infinitesimal model where variation is lost by genetic drift and gained by mutation and therefore the minimum population size to maintain genetic variation is a function of mutation rate. For genes with effects
Genetic materials at the gene engineering division, RIKEN BioResource Center.
Yokoyama, Kazunari K; Murata, Takehide; Pan, Jianzhi; Nakade, Koji; Kishikawa, Shotaro; Ugai, Hideyo; Kimura, Makoto; Kujime, Yukari; Hirose, Megumi; Masuzaki, Satoko; Yamasaki, Takahito; Kurihara, Chitose; Okubo, Masato; Nakano, Yuri; Kusa, Yuka; Yoshikawa, Akiko; Inabe, Kumiko; Ueno, Kazuko; Obata, Yuichi
Genetic materials are one of the most important and fundamental research resources for studying biological phenomena. Scientific need for genetic materials has been increasing and will never cease. Ever since it was established as RIKEN DNA Bank in 1987, the Gene Engineering Division of RIKEN BioResource Center (BRC) has been engaged in the collection, maintenance, storage, propagation, quality control, and distribution of genetic resources developed mainly by the Japanese research community. When RIKEN BRC was inaugurated in 2001, RIKEN DNA Bank was incorporated as one of its six Divisions, the Gene Engineering Division. The Gene Engineering Division was selected as a core facility for the genetic resources of mammalian and microbe origin by the National BioResource Project (NBRP) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan in 2002. With support from the scientific community, the Division now holds over 3 million clones of genetic materials for distribution. The genetic resources include cloned DNAs, gene libraries (e.g., cDNA and genomic DNA cloned into phage, cosmid, BAC, phosmid, and YAC), vectors, hosts, recombinant viruses, and ordered library sets derived from animal cells, including human and mouse cells, microorganisms, and viruses. Recently genetic materials produced by a few MEXT national research projects were transferred to the Gene Engineering Division for further dissemination. The Gene Engineering Division performs rigorous quality control of reproducibility, restriction enzyme mapping and nucleotide sequences of clones to ensure the reproducibility of in vivo and in vitro experiments. Users can easily access our genetic materials through the internet and obtain the DNA resources for a minimal fee. Not only the materials, but also information of features and technology related to the materials are provided via the web site of RIKEN BRC. Training courses are also given to transfer the technology for handling viral vectors. RIKEN BRC supports scientists around the world in the use of valuable genetic materials. PMID:20484845
Genetic engineering of industrial strains of Saccharomyces cerevisiae.
Le Borgne, Sylvie
Genetic engineering has been successfully applied to Saccharomyces cerevisiae laboratory strains for different purposes: extension of substrate range, improvement of productivity and yield, elimination of by-products, improvement of process performance and cellular properties, and extension of product range. The potential of genetically engineered yeasts for the massive production of biofuels as bioethanol and other nonfuel products from renewable resources as lignocellulosic biomass hydrolysates has been recognized. For such applications, robust industrial strains of S. cerevisiae have to be used. Here, some relevant genetic and genomic characteristics of industrial strains are discussed in relation to the problematic of the genetic engineering of such strains. General molecular tools applicable to the manipulation of S. cerevisiae industrial strains are presented and examples of genetically engineered industrial strains developed for the production of bioethanol from lignocellulosic biomass are given. PMID:22160914
Genetic Engineering and the Amelioration of Genetic Defect
ERIC Educational Resources Information Center
Lederberg, Joshua
Discusses the claims for a brave new world of genetic manipulation" and concludes that if we could agree upon applying genetic (or any other effective) remedies to global problems we probably would need no rescourse to them. Suggests that effective methods of preventing genetic disease are prevention of mutations and detection and containment ofģ
Possible people, complaints, and the distinction between genetic planning and genetic engineering
James J Delaney
Advances in the understanding of genetics have led to the belief that it may become possible to use genetic engineering to manipulate the DNA of humans at the embryonic stage to produce certain desirable traits. Although this currently cannot be done on a large scale, many people nevertheless object in principle to such practices. Most often, they argue that genetic
Biomimetic Evolutionary Reverse Engineering of Genetic Regulatory Networks
Daniel Marbach; Claudio Mattiussi; Dario Floreano
The eective reverse engineering of biochemical networks is one of the great challenges of systems biology. The contribution of this paper is two-fold: 1) We introduce a new method for reverse engineering genetic regulatory networks from gene expression data; 2) We demon- strate how nonlinear gene networks can be inferred from steady-state data alone. The reverse engineering method is based
Genetically Engineered Mouse Models for Drug Development and Preclinical Trials
Lee, Ho
Drug development and preclinical trials are challenging processes and more than 80% to 90% of drug candidates fail to gain approval from the United States Food and Drug Administration. Predictive and efficient tools are required to discover high quality targets and increase the probability of success in the process of new drug development. One such solution to the challenges faced in the development of new drugs and combination therapies is the use of low-cost and experimentally manageable in vivo animal models. Since the 1980Æs, scientists have been able to genetically modify the mouse genome by removing or replacing a specific gene, which has improved the identification and validation of target genes of interest. Now genetically engineered mouse models (GEMMs) are widely used and have proved to be a powerful tool in drug discovery processes. This review particularly covers recent fascinating technologies for drug discovery and preclinical trials, targeted transgenesis and RNAi mouse, including application and combination of inducible system. Improvements in technologies and the development of new GEMMs are expected to guide future applications of these models to drug discovery and preclinical trials. PMID:25143803
Seeking Perfection: A Kantian Look at Human Genetic Engineering
Martin Gunderson
It is tempting to argue that Kantian moral philosophy justifies prohibiting both human germ-line genetic engineering and non-therapeutic\\u000a genetic engineering because they fail to respect human dignity. There are, however, good reasons for resisting this temptation.\\u000a In fact, KantÆs moral philosophy provides reasons that support genetic engineeringŚeven germ-line and non-therapeutic. This\\u000a is true of KantÆs imperfect duties to seek oneÆs
Modeling the Diagnostic Criteria for Alcohol Dependence with Genetic Animal Models
Kendler, Kenneth S.; Hitzemann, Robert J.
A diagnosis of alcohol dependence (AD) using the DSM-IV-R is categorical, based on an individualÆs manifestation of three or more symptoms from a list of seven. AD risk can be traced to both genetic and environmental sources. Most genetic studies of AD risk implicitly assume that an AD diagnosis represents a single underlying genetic factor. We recently found that the criteria for an AD diagnosis represent three somewhat distinct genetic paths to individual risk. Specifically, heavy use and tolerance versus withdrawal and continued use despite problems reflected separate genetic factors. However, some data suggest that genetic risk for AD is adequately described with a single underlying genetic risk factor. Rodent animal models for alcohol-related phenotypes typically target discrete aspects of the complex human AD diagnosis. Here, we review the literature derived from genetic animal models in an attempt to determine whether they support a single-factor or multiple-factor genetic structure. We conclude that there is modest support in the animal literature that alcohol tolerance and withdrawal reflect distinct genetic risk factors, in agreement with our human data. We suggest areas where more research could clarify this attempt to align the rodent and human data. PMID:21910077
Congenital heart diseases in small animals: Part I. Genetic pathways and potential candidate genes
Changbaig Hyun; Lopeti Lavulo
Proper cardiac morphogenesis requires a series of specific cell and tissue interactions driven by several cardiac transcription factors and downstream cardiac genes. To date, a number of genetic aetiologies responsible for human congenital heart defects (CHDs) have been identified, although none has been found for CHDs in small animals. Most gene mutations responsible for human CHDs exist in genetic pathways
Colonization genetics of an animal-dispersed plant (Vaccinium membranaceum) at Mount St Helens, Washington
Population founding and spatial spread may profoundly influence later population genetic structure, but their effects are difficult to quantify when population history is unknown. We examined the genetic effects of founder group formation in a recently founded population of the animal-dispersed Vaccinium membranaceum (black huckleberry) on new volcanic deposits at Mount St Helens (Washington, USA) 24 years post-eruption. Using amplified
Genetic diversity of Clostridium perfringens type A isolates from animals, food poisoning outbreaks and sludge
Anders Johansson; Anna Aspan; Elisabeth Bagge; Viveca BÕverud; Bj÷rn E Engstr÷m; Karl-Erik Johansson
BACKGROUND: Clostridium perfringens, a serious pathogen, causes enteric diseases in domestic animals and food poisoning in humans. The epidemiological relationship between C. perfringens isolates from the same source has previously been investigated chiefly by pulsed-field gel electrophoresis (PFGE). In this study the genetic diversity of C. perfringens isolated from various animals, from food poisoning outbreaks and from sludge was investigated.
Genetically engineered phage fibers and coatings for antibacterial applications
Mao, Joan Y
Multifunctionality can be imparted to protein-based fibers and coatings via either synthetic or biological approaches. Here, we demonstrate potent antimicrobial functionality of genetically engineered, phage-based fibers ...
Concepts and methods were identified for their utility in evaluating the persistence and potential perturbations of genetically engineered microorganisms in the environment. Novel uses of DNA reassociation kinetics and gene probe technologies, in conjunction with conventional bac...
Perspective on Models in Theoretical and Practical Traditions of Knowledge: The Example of Otto Engine Animations
ERIC Educational Resources Information Center
Haglund, Jesper; Stromdahl, Helge
Nineteen informants (n = 19) were asked to study and comment two computer animations of the Otto combustion engine. One animation was non-interactive and realistic in the sense of depicting a physical engine. The other animation was more idealised, interactive and synchronised with a dynamic PV-graph. The informants represented practical andģ
Genetic engineering of doxorubicin production in Streptomyces peucetius : a review
C R Hutchinson; A L Colombo
ĀĀThe genetics and biochemistry of daunorubicin and doxorubicin production by Streptomyces peucetius is reviewed, with a focus on how such information can be used for the genetic engineering of strains having improved titers\\u000a of these two antitumor antibiotics.
Genetic engineering and stem cells: combinatorial approaches for cardiac cell therapy [Cellular\\/Tissue Engineering
Robert D. Kirkton; Nenad Bursac
The use of genetic engineering to induce or alter specific protein expression in stem cells has already facilitated research in this field and may, additionally, offer a potential route for designing more efficient cell sources for cardiac repair. As the feasibility of stem cell genetic manipulation has already been proved and as genetic techniques continue to advance, the combinatorial approach
Illuminating Cancer Systems With Genetically-Engineered Mouse Models and Coupled Luciferase Reporters In Vivo
Kocher, Brandon; Piwnica-Worms, David
Bioluminescent imaging (BLI) is a powerful non-invasive tool that has dramatically accelerated the in vivo interrogation of cancer systems and longitudinal analysis of mouse models of cancer over the past decade. Various luciferase enzymes have been genetically engineered into mouse models (GEMMs) of cancer which permit investigation of cellular and molecular events associated with oncogenic transcription, post-transcriptional processing, protein-protein interactions, transformation and oncogene addiction in live cells and animals. Luciferase-coupled GEMMs ultimately serve as a non-invasive, repetitive, longitudinal, and physiological means by which cancer systems and therapeutic responses can be investigated accurately within the autochthonous context of a living animal. PMID:23585416
Promoting Student Engagement in the Animal Sciences: an "Academic Pedigree" Project. An Academic Pedigree project was incorporated into the junior-level Animal Breeding and Genetics
Grissino-Mayer, Henri D.
engagement of students in a genetics course. Groups of students were assigned to trace a faculty member in Animal Science, and increase general engagement of students in a genetics course, an "Academic PedigreePromoting Student Engagement in the Animal Sciences: an "Academic Pedigree" Project. CJ Kojima
Fernandez, Thomas
REVERSE ENGINEERING AND AUTOMATIC SYNTHESIS OF METABOLIC PATHWAYS FROM OBSERVED DATA USING GENETIC of Medicine Department of Electrical Engineering Stanford University, Stanford, California, koza (reverse engineer) a network of chemical reactions from observed time-domain data. Genetic programming
Chloroplast Genetic Engineering: Recent Advances and Future Perspectives
Justin James Grevich; Henry Daniell
Chloroplast genetic engineering offers a number of unique advantages, including a high-level of transgene expression, multi-gene engineering in a single transformation event, transgene containment via maternal inheritance, lack of gene silencing, position and pleiotropic effects, and undesirable foreign DNA. Thus far, over forty transgenes have been stably integrated and expressed via the tobacco chloroplast genome to confer important agronomic traits,
Chapter VIII. Contributions of propagation techniques and genetic modification to breeding - genetic engineering for disease resistance
Technology Transfer Automated Retrieval System (TEKTRAN)
Genetic engineering offers an opportunity to develop flower bulb crops with resistance to fungal, viral, and bacterial pathogens. Several of the flower bulb crops, Lilium spp., Gladiolus, Zantedeschia, Muscari, Hyacinthus, Narcissus, Ornithogalum, Iris, and Alstroemeria, have been transformed with t...
SELECTION FOR GROWTH RATE AND POSSIBILITIES FOR GENETIC CHANGE IN ATLANTIC SALMON Dep. of Animal Genetics and Bveeding, Agvicultuval University of Norway,
Paris-Sud XI, Universitķ de
SELECTION FOR GROWTH RATE AND POSSIBILITIES FOR GENETIC CHANGE IN ATLANTIC SALMON T. GJEDREM Dep. of Animal Genetics and Bveeding, Agvicultuval University of Norway, I432 ┼S- NLH, Norway The possibility for genetic change in production traits of salmon looks good when consider- ing the magnitude of genetic
Genetic and somatic effects in animals maintained on tritiated water
Carsten, A.L.; Brooks, A.; Commerford, S.L.; Cronkite, E.P.
The possible genetic (dominant lethal mutations (DLM) and cytogenetic changes in the regenerating liver) and somatic (hematopoietic stem cell changes, growth and nonspecific life time shortening) effects in mice maintained on tritiated water (HTO) over two generations was investigated. Results to date are summarized. (ACR)
Genetic Animal Models of Alcohol and Drug Abuse
John C. Crabbe; John K. Belknap; Kari J. Buck
Behavioral and pharmacological responses of selectively bred and inbred rodent lines have been analyzed to elucidate many features of drug sensitivity and the adverse effects of drugs, the underlying mechanisms of drug tolerance and dependence, and the motivational states underlying drug reward and aversion. Genetic mapping of quantitative trait loci (QTLs) has been used to identify provisional chromosomal locations of
Distribution of Genetic Marker Concentrations for Fecal Indicator Bacteria in Sewage and Animal Feces
Kelty, Catherine A.; Varma, Manju; Sivaganesan, Mano; Haugland, Richard A.
Very little is known about the density and distribution of fecal indicator bacteria (FIB) genetic markers measured by quantitative real-time PCR (qPCR) in fecal pollution sources. Before qPCR-based FIB technologies can be applied to waste management and public health risk applications, it is vital to characterize the concentrations of these genetic markers in pollution sources (i.e., untreated wastewater and animal feces). We report the distribution of rRNA genetic markers for several general FIB groups, including Clostridium spp., Escherichia coli, enterococci, and Bacteroidales, as determined by qPCR on reference collections consisting of 54 primary influent sewage samples collected from treatment facilities across the United States and fecal samples representing 20 different animal species. Based on raw sewage sample collection data, individual FIB genetic markers exhibited a remarkable similarity in concentration estimates from locations across the United States ranging from Hawaii to Florida. However, there was no significant correlation between genetic markers for most FIB combinations (P > 0.05). In addition, large differences (up to 5 log10 copies) in the abundance of FIB genetic markers were observed between animal species, emphasizing the importance of indicator microorganism selection and animal source contribution for future FIB applications. PMID:22504809
Genetic manipulation of sex differentiation and phenotype in domestic animals
D. W. Silversides; N. Pilon; R. Behdjani; A. Boyer; I. Daneau; J. Lussier
In mammals, a gene based sex determination system ensures that approximately 50% of offspring will be of the male sex and 50% will be of the female sex. In domestic animal production systems, this ratio is not always ideal. Recent advances in our understanding of the molecular biology of sex determination and differentiation, as well as in the control of
Use of Genetically Modified Viruses and Genetically Engineered Virus-vector Vaccines: Environmental Effects
Vivian S. W. Chan
Despite major therapeutic advances, infectious diseases remain highly problematic. Recent advancements in technology in producing DNA-based vaccines, together with the growing knowledge of the immune system, have provided new insights into the identification of the epitopes needed to target the development of highly targeted vaccines. Genetically modified (GM) viruses and genetically engineered virus-vector vaccines possess significant unpredictability and a number
Synopsis - Developments in in vitro fertilization techniques and recombinant DNA technology are improving the technical feasibility of genetically manipulating human embryos. The combinationof these technologies allows a new form of eugenic selection to be practiced and some IVF practitioners and researchers are advocating that genetic disorders can be eradicated from future generations in the human population. Prevention of the
Prospects for Genetic Engineering in Plants
Technology Transfer Automated Retrieval System (TEKTRAN)
Genetically modified plants now constitute a significant portion of the worlds agricultural output. Genetically modified corn, soybean, canola, rice, and cotton are being adopted by growers in both industrialized and developing nations at an increasing rate. The most popular products have been eng...
The use of whole food animal studies in the safety assessment of genetically modified crops: limitations and recommendations.
Bartholomaeus, Andrew; Parrott, Wayne; Bondy, Genevieve; Walker, Kate
There is disagreement internationally across major regulatory jurisdictions on the relevance and utility of whole food (WF) toxicity studies on GM crops, with no harmonization of data or regulatory requirements. The scientific value, and therefore animal ethics, of WF studies on GM crops is a matter addressable from the wealth of data available on commercialized GM crops and WF studies on irradiated foods. We reviewed available GM crop WF studies and considered the extent to which they add to the information from agronomic and compositional analyses. No WF toxicity study was identified that convincingly demonstrated toxicological concern or that called into question the adequacy, sufficiency, and reliability of safety assessments based on crop molecular characterization, transgene source, agronomic characteristics, and/or compositional analysis of the GM crop and its near-isogenic line. Predictions of safety based on crop genetics and compositional analyses have provided complete concordance with the results of well-conducted animal testing. However, this concordance is primarily due to the improbability of de novo generation of toxic substances in crop plants using genetic engineering practices and due to the weakness of WF toxicity studies in general. Thus, based on the comparative robustness and reliability of compositional and agronomic considerations and on the absence of any scientific basis for a significant potential for de novo generation of toxicologically significant compositional alterations as a sole result of transgene insertion, the conclusion of this review is that WF animal toxicity studies are unnecessary and scientifically unjustifiable. PMID:24164514
The use of whole food animal studies in the safety assessment of genetically modified crops: Limitations and recommendations
Bartholomaeus, Andrew; Parrott, Wayne; Bondy, Genevieve
There is disagreement internationally across major regulatory jurisdictions on the relevance and utility of whole food (WF) toxicity studies on GM crops, with no harmonization of data or regulatory requirements. The scientific value, and therefore animal ethics, of WF studies on GM crops is a matter addressable from the wealth of data available on commercialized GM crops and WF studies on irradiated foods. We reviewed available GM crop WF studies and considered the extent to which they add to the information from agronomic and compositional analyses. No WF toxicity study was identified that convincingly demonstrated toxicological concern or that called into question the adequacy, sufficiency, and reliability of safety assessments based on crop molecular characterization, transgene source, agronomic characteristics, and/or compositional analysis of the GM crop and its near-isogenic line. Predictions of safety based on crop genetics and compositional analyses have provided complete concordance with the results of well-conducted animal testing. However, this concordance is primarily due to the improbability of de novo generation of toxic substances in crop plants using genetic engineering practices and due to the weakness of WF toxicity studies in general. Thus, based on the comparative robustness and reliability of compositional and agronomic considerations and on the absence of any scientific basis for a significant potential for de novo generation of toxicologically significant compositional alterations as a sole result of transgene insertion, the conclusion of this review is that WF animal toxicity studies are unnecessary and scientifically unjustifiable. PMID:24164514
Application of multiple trait analysis in animal breeding research Department of Animal Breeding and Genetics
Paris-Sud XI, Universitķ de
The free fatty acid (FFA) content in milk from 60 cows were determined. Fresh and stored samples from both Breeding and Genetics Swedish University of Agricultural Sciences, S-750 07 Uppsala, Sweden lifetime milk production, productive life, level of production and a lifetime profit function in cows A
The Animal Genetic Resource Information Network (AnimalGRIN) Database: A Database Design & Implementation Case
ERIC Educational Resources Information Center
Irwin, Gretchen; Wessel, Lark; Blackman, Harvey
This case describes a database redesign project for the United States Department of Agriculture's National Animal Germplasm Program (NAGP). The case provides a valuable context for teaching and practicing database analysis, design, and implementation skills, and can be used as the basis for a semester-long team project. The case demonstrates theģ
Regulatory Oversight of Genetically Engineered Microorganisms: Has Regulation Inhibited Innovation?
Wrubel; Krimsky; Anderson
/ Using detailed interviews with company representatives and researchers in the field, this paper examines the factors that might account for the slow pace of development of genetically engineered microorganisms (GEMs) intended for environmental release. We specifically analyzed the role of the regulatory system in shaping innovation. We identified at least two cases where industry decided to discontinue the development of a genetically engineered microbial product because of concerns over regulatory oversight. However, most often industry decisions to continue or halt development of GEMs were based on an evaluation of the particular product's efficacy and potential for profitability. Thus the inability of GEMs to perform up to expectations in the field, rather than the regulatory constraints, appears to be the factor responsible for the slow pace of development. KEY WORDS: Genetically engineered microorganisms; Biotechnology; Regulation of biotechnology; Innovation; Environmental release PMID:9175544
Regulatory steps associated with use of value-added recombinant proteins and peptides screened in high-throughput for expression in genetically engineered starch and cellulosic fuel ethanol yeast strains
Technology Transfer Automated Retrieval System (TEKTRAN)
Recombinant proteins expressed in animals have been a public concern as a perceived risk to the consumer. Animals are currently being treated with genetically engineered biologicals, such as growth hormone, or fed genetically modified plants. Similarly, various commercially-valuable proteins or pe...
Genetic structure and diversity of animal populations exposed to metal pollution.
Mussali-Galante, Patricia; Tovar-Sßnchez, EfraĒn; Valverde, Mahara; Rojas, Emilio
Studying the genetic diversity of wild populations that are affected by pollution provides a basis for estimating the risks of environmental contamination to both wildlife, and indirectly to humans. Such research strives to produce both a better understanding of the underlying mechanisms by which genetic diversity is affected,and the long-term effects of the pollutants involved.In this review, we summarize key aspects of the field of genetic ecotoxicology that encompasses using genetic patterns to examine metal pollutants as environmental stressors of natural animal populations. We address genetic changes that result from xenobiotic exposure versus genetic alterations that result from natural ecological processes. We also describe the relationship between metal exposure and changes in the genetic diversity of chronically exposed populations, and how the affected populations respond to environmental stress. Further, we assess the genetic diversity of animal populations that were exposed to metals, focusing on the literature that has been published since the year 2000.Our review disclosed that the most common metals found in aquatic and terrestrial ecosystems were Cd, Zn, Cu and Pb; however, differences in the occurrence between aquatic (Cd=Zn>Cu>Pb>Hg) and terrestrial (Cu>Cd>Pb>Zn>Ni)environments were observed. Several molecular markers were used to assess genetic diversity in impacted populations, the order of the most common ones of which were SSR's > allozyme > RAPD's > mtDNA sequencing> other molecular markers.Genetic diversity was reduced for nearly all animal populations that were exposed to a single metal, or a mixture of metals in aquatic ecosystems (except in Hyalella azteca, Littorina littorea, Salmo trutta, and Gobio gobio); however, the pattern was less clear when terrestrial ecosystems were analyzed.We propose that future research in the topic area of this paper emphasizes seven key areas of activity that pertain to the methodological design of genetic ecotoxicological studies. Collectively, these points are designed to provide more accurate data and a deeper understanding of the relationship between alterations in genetic diversity of impacted populations and metal exposures. In particular, we believe that the exact nature of all tested chemical pollutants be clearly described, biomarkers be included, sentinel organisms be used, testing be performed at multiple experimental sites, reference populations be sampled in close geographical proximity to where pollution occurs, and genetic structure parameters and high-throughput technology be more actively employed. Furthermore, we propose a new class of biomarkers,termed "biomarkers of permanent effect," which may include measures of genetic variability in impacted populations. PMID:24158580
ABE Agricultural and Biological Engineering F9 ADDL Animal Disease Diagnostic Lab G10
ABE Agricultural and Biological Engineering F9 ADDL Animal Disease Diagnostic Lab G10 AERO Aerospace Science Laboratory C11 AGAD Agricultural Administration Building G8 AHF Animal Holding Facility G10 AQUA Boilermaker Aquatic Center D6 AR Armory G6 ARMS Armstrong (Neil) Hall of Engineering G5 ASTL
Conceptual, methodological and ethical issues in genetic engineering (1989)
A. Falek
An introduction to the potential of gene therapy to alleviate illness and death particularly for many rare human genetic disorders\\u000a and specific forms of cancer is presented. At present, genetic engineering, that is gene therapy to correct some of these\\u000a disorders based on new molecular biology procedures is a possibility in the near future especially those with single gene\\u000a mendelian
Adoptive immunotherapy of cancer utilizing genetically engineered lymphocytes.
Ikeda, Hiroaki; Shiku, Hiroshi
It is becoming increasingly clear that adoptive immunotherapy with genetically engineered T cells has the potential to control and even cure cancer in some patients. On the other hand, severe adverse events associated with efficacy have frequently been reported in clinical trials. Current and near-future challenges for the development of adoptive immunotherapy of cancer using genetically engineered T cells include minimization and prediction of adverse events; identification of new and effective targets, including patient-specific mutations; improvement in T cell functionality, persistence, and memory formation capacity; and utilization of allogeneic or cell line-based T cells. PMID:26041411
Current development in genetic engineering strategies of Bacillus species
The complete sequencing and annotation of the genomes of industrially-important Bacillus species has enhanced our understanding of their properties, and allowed advances in genetic manipulations in other Bacillus species. Post-genomic studies require simple and highly efficient tools to enable genetic manipulation. Here, we summarize the recent progress in genetic engineering strategies for Bacillus species. We review the available genetic tools that have been developed in Bacillus species, as well as methods developed in other species that may also be applicable in Bacillus. Furthermore, we address the limitations and challenges of the existing methods, and discuss the future research prospects in developing novel and useful tools for genetic modification of Bacillus species. PMID:24885003
Journal of Assisted Reproduction and Genetics, Vol. 18, No. 5, 2001 ANIMAL EXPERIMENTATION
Kim, Daesoo
Journal of Assisted Reproduction and Genetics, Vol. 18, No. 5, 2001 ANIMAL EXPERIMENTATION, Fertilization, and Embryo Development1 DOOSEOK CHOI,2,4 EUNYOUNG LEE,2 SEONGSOO HWANG,2 KISUN JUN,3 DAESOO KIM,3 mutation in mouse sperm in the acrosome reaction, fertilization, and embryo de- velopment. Methods: Study
Diverse Plant and Animal Genetic Records from Holocene and Pleistocene Sediments
Eske Willerslev; Anders J. Hansen; Jonas Binladen; Tina B. Brand; M. Thomas P. Gilbert; Beth Shapiro; Michael Bunce; Carsten Wiuf; David A. Gilichinsky; Alan Cooper
Genetic analyses of permafrost and temperate sediments reveal that plant and animal DNA may be preserved for long periods, even in the absence of obvious macrofossils. In Siberia, five permafrost cores ranging from 400,000 to 10,000 years old contained at least 19 different plant taxa, including the oldest authenticated ancient DNA sequences known, and megafaunal sequences including mammoth, bison, and
Withdrawal emotional-regulation in infant rats from genetic animal models of depression
Y. Braw; O. Malkesman; A. Merenlender; A. Bercovich; M. Dagan; D. H. Overstreet; A. Weller
Children of depressed parents exhibit high rates of emotion-dysregulation, characterized by excessive withdrawal or approach strategies toward the mother in infancy. The understanding of factors affecting the establishment of these behavioral deficits is limited. The current study utilized two genetic animal models of depression, the Wistar¢Kyoto (WKY) and Flinders Sensitive Line (FSL) rat strains. In addition, in order to assess
Assessing the Transfer of Genetically Modified DNA from Feed to Animal Tissues
Raffaele Mazza; Mirko Soave; Mauro Morlacchini; Gianfranco Piva; Adriano Marocco
In Europe, public and scientific concerns about the environmental and food safety of GM (Genetically Modified) crops overshadow the potential benefits offered by crop biotechnology to improve food quality. One of the concerns regarding the use of GM food in human and animal nutrition is the effect that newly introduced sequences may have on the organism. In this paper, we
A review on SNP and other types of molecular markers and their use in animal genetics
Alain Vignal; Denis Milan; Magali SanCristobal; Andrķ Eggen
During the last ten years, the use of molecular markers, revealing polymorphism at the DNA level, has been playing an increasing part in animal genetics studies. Amongst others, the microsatellite DNA marker has been the most widely used, due to its easy use by simple PCR, followed by a denaturing gel electrophoresis for allele size determination, and to the high
Detection and quantification of genetically engineered crops
Asfaw Adugna; Tewodros Mesfin
Summary Genetically modified organisms (GMOs) have recently attracted the attention of agricultural, medical and food scientists and governments of many countries in the world due to an increasing concern that the recombinant gene(s) inserted into an organism may result in unforeseen effects. Therefore, there is a need to regulate each transgenic event so that the officially approved events will be
Neuropathology and Animal Models of Autism: Genetic and Environmental Factors
Gadad, Bharathi S.; Young, Keith A.; German, Dwight C.
Autism is a heterogeneous behaviorally defined neurodevelopmental disorder. It is defined by the presence of marked social deficits, specific language abnormalities, and stereotyped repetitive patterns of behavior. Because of the variability in the behavioral phenotype of the disorder among patients, the term autism spectrum disorder has been established. In the first part of this review, we provide an overview of neuropathological findings from studies of autism postmortem brains and identify the cerebellum as one of the key brain regions that can play a role in the autism phenotype. We review research findings that indicate possible links between the environment and autism including the role of mercury and immune-related factors. Because both genes and environment can alter the structure of the developing brain in different ways, it is not surprising that there is heterogeneity in the behavioral and neuropathological phenotypes of autism spectrum disorders. Finally, we describe animal models of autism that occur following insertion of different autism-related genes and exposure to environmental factors, highlighting those models which exhibit both autism-like behavior and neuropathology. PMID:24151553
Technology Transfer Automated Retrieval System (TEKTRAN)
Mastitis is a $2 billion a year problem for the dairy industry. Neither the animals immune system nor conventional prophylactic strategies have been able to effectively address the problem. We propose using new biotechnology tools to introduce genes into cattle from other species to solve this pro...
Biomechanical considerations of animal models used in tissue engineering of bone
Michael A. K. Liebschner
Tissue engineering combines the aspects of cell biology, engineering, material science, and surgery to generate new functional tissue, and provides an important approach to the repair of segmental defects and in restoring biomechanical function. The development of tissue-engineering strategies into clinical therapeutic protocols requires extensive, preclinical experimentation in appropriate animal models. The ultimate success of any treatment strategy must be
Technology Transfer Automated Retrieval System (TEKTRAN)
Through genetic engineering of peanut, we have focused mainly on two levels of protection against aflatoxin contamination: the entry of spores through insect-damaged tissues and the growth of the fungus after entry, although interfering with the aflatoxin biosynthetic pathway also is of interest. T...
A Simple Interactive Introduction to Teaching Genetic Engineering
ERIC Educational Resources Information Center
Child, Paula
In the UK, at key stage 4, students aged 14-15 studying GCSE Core Science or Unit 1 of the GCSE Biology course are required to be able to describe the process of genetic engineering to produce bacteria that can produce insulin. The simple interactive introduction described in this article allows students to consider the problem, devise a model andģ
Environmental risks and fate of genetically engineered microorganisms in soil
J. D. van Elsas; J. T. Trevors
Genetically engineered microorganisms in the environment will prove to be useful if they provide a means by which to perform a desirable function in the environment, whilst not posing unacceptable risks to any component of the environment. Moreover, the uncertainty associated with the environmental release of living organisms due to the high density of cells that will be released and
Harnessing genetically engineered mouse models for preclinical testing
Ana I. Robles; Lyuba Varticovski
Recent studies cast doubt on the value of traditionally used models as tools for testing therapies for human cancer. Although the standard practice of xenografting tumors into immunocompromised mice generates reproducible tumors, drug testing in these models has low predictive power when compared to the clinical responses in Phase II trials. The use of tumor-bearing genetically engineered mouse models holds
Genetically Engineered Mice: Tools To Understand Craniofacial Development
Michael A. Ignelzi; Yi-Hsin Liu; Robert E. Maxson; Malcolm L. Snead
In this review, we provide a survey of the experimental approaches used to generate genetically engineered mice. Two specific examples are presented that demonstrate the applicability of these approaches to craniofacial development. In the first, a promoter analysis of the Msx2 gene is presented which illustrates the cis regulatory interactions that define cell-specific gene expression. In the second, a mouse
Dep. Of Biotechnology & Genetic Engineering Study plan(2005-2006)
Dep. Of Biotechnology & Genetic Engineering Study plan(2005-2006) First semester or Con 212241Analytical Chemistry3212103240281 Introduction to Biotechnology2 240107+1 30102 2122423240233240462Bioinformatics2 240231+ 240335 240338Immunology lab1 240337 or Con 240322Plant Biotechnology3240281
Genetically Engineered Materials for Biofuels Production
NASA Astrophysics Data System (ADS)
Raab, Michael
Agrivida, Inc., is an agricultural biotechnology company developing industrial crop feedstocks for the fuel and chemical industries. Agrivida's crops have improved processing traits that enable efficient, low cost conversion of the crops' cellulosic components into fermentable sugars. Currently, pretreatment and enzymatic conversion of the major cell wall components, cellulose and hemicellulose, into fermentable sugars is the most expensive processing step that prevents widespread adoption of biomass in biofuels processes. To lower production costs we are consolidating pretreatment and enzyme production within the crop. In this strategy, transgenic plants express engineered cell wall degrading enzymes in an inactive form, which can be reactivated after harvest. We have engineered protein elements that disrupt enzyme activity during normal plant growth. Upon exposure to specific processing conditions, the engineered enzymes are converted into their active forms. This mechanism significantly lowers pretreatment costs and enzyme loadings (>75% reduction) below those currently available to the industry.
Animal Genetic Resource Trade Flows: The Utilization of Newly Imported Breeds and the Gene Flow of Imported Animals in the United States of America
Technology Transfer Automated Retrieval System (TEKTRAN)
Animal germplasm exchange has recently received attention as a product of the FAOÆs State of the WorldÆs Animal Genetic Resources effort. Some have advocated a need to explore policies and regulations on the exchange of germplasm. However, there has been little comprehensive assessment of either th...
Genetic engineering possibilities for CELSS: A bibliography and summary of techniques
NASA Technical Reports Server (NTRS)
Johnson, E. J.
A bibliography of the most useful techniques employed in genetic engineering of higher plants, bacteria associated with plants, and plant cell cultures is provided. A resume of state-of-the-art genetic engineering of plants and bacteria is presented. The potential application of plant bacterial genetic engineering to CELSS (Controlled Ecological Life Support System) program and future research needs are discussed.
Attitudes towards genetic engineering between change and stability: Results of a panel study
Dieter Urban; Uwe Pfenning
Genetic engineering is not only a new modern technology but it is also a social object of technological assessment research. Genetic engineering is associated with moral and ethical concerns in society, political decisions, ecological impacts, and economics. These impacts are the foundation for the social linkages of genetic engineering. For that social linkages and cognitive association we have chosen the
The ecology and evolution of animal medication: genetically fixed response versus phenotypic plasticity.
Choisy, Marc; de Roode, Jacobus C
Animal medication against parasites can occur either as a genetically fixed (constitutive) or phenotypically plastic (induced) behavior. Taking the tritrophic interaction between the monarch butterfly Danaus plexippus, its protozoan parasite Ophryocystis elektroscirrha, and its food plant Asclepias spp. as a test case, we develop a game-theory model to identify the epidemiological (parasite prevalence and virulence) and environmental (plant toxicity and abundance) conditions that predict the evolution of genetically fixed versus phenotypically plastic forms of medication. Our model shows that the relative benefits (the antiparasitic properties of medicinal food) and costs (side effects of medicine, the costs of searching for medicine, and the costs of plasticity itself) crucially determine whether medication is genetically fixed or phenotypically plastic. Our model suggests that animals evolve phenotypic plasticity when parasite risk (a combination of virulence and prevalence and thus a measure of the strength of parasite-mediated selection) is relatively low to moderately high and genetically fixed medication when parasite risk becomes very high. The latter occurs because at high parasite risk, the costs of plasticity are outweighed by the benefits of medication. Our model provides a simple and general framework to study the conditions that drive the evolution of alternative forms of animal medication. PMID:25061676
Advances in genetic modification of farm animals using zinc-finger nucleases (ZFN).
Petersen, Bjoern; Niemann, Heiner
Genome editing tools (GET), including zinc-finger nucleases (ZFN), transcription activator-like endonucleases (TALENS), and meganucleases possess long recognition sites and are thus capable of cutting DNA in a very specific manner. These genome editing tools mediate targeted genetic alterations by enhancing DNA mutation frequency via induction of double-strand breaks at a predetermined genomic site. Compared to conventional homologous recombination based gene targeting, GETs can increase gene targeting and gene disruption via mutagenic DNA repair more than 10,000-fold. Recently, a novel class of genome editing tools was described that uses RNAs to target a specific genomic site. The CRISPR/Cas9 system is capable of targeting even multiple genomic sites in one shot and thus could be superior to ZFNs or TALEN. Current results indicate that these tools can be successfully employed in a broad range of organisms which renders them useful for improving the understanding of complex physiological systems, producing transgenic animals, including creating large animal models for human diseases, creating specific cell lines, and plants, and even for treating human genetic diseases. This review provides an update on the use of ZFNs to modify the genome of farm animals, summarizes current knowledge on the underlying mechanism, and discusses new opportunities for generating genetically modified farm animals. PMID:25596823
Between creation, evolution and genetic engineering : biology in need of a new bioethics?
J. A. Gupta
Technological interventions into biological\\u000aprocesses through genetic engineering in\\u000athe twenty-fi rst century could speed up evolution\\u000aat the velocity of light years in comparison\\u000awith the millions of years it took for\\u000aHomo sapiens to reach this stage of evolution\\u000auntil this new millennium. Will these\\u000atechnological assays lead to an eventual recrafting\\u000aof animal and human biology and
Developing a systematic strategy incorporating ethical, animal welfare and practical principles to guide the genetic improvement of dairy cattle
MW Fisher; DJ Mellor
People have complex and diverse relationships and interactions with, and expectations of, animals; relationships which are very important. In making sense of this complexity, we draw on our values. The objective of this study was to reflect upon, develop and articulate key values guiding the genetic improvement of dairy cattle.Animal husbandry is guided by the philosophy that while animals serve
Colonization genetics of an animal-dispersed plant (Vaccinium membranaceum) at Mount St Helens, Washington.
Yang, S; Bishop, J G; Webster, M S
Population founding and spatial spread may profoundly influence later population genetic structure, but their effects are difficult to quantify when population history is unknown. We examined the genetic effects of founder group formation in a recently founded population of the animal-dispersed Vaccinium membranaceum (black huckleberry) on new volcanic deposits at Mount St Helens (Washington, USA) 24 years post-eruption. Using amplified fragment length polymorphisms and assignment tests, we determined sources of the newly founded population and characterized genetic variation within new and source populations. Our analyses indicate that while founders were derived from many sources, about half originated from a small number of plants that survived the 1980 eruption in pockets of remnant soil embedded within primary successional areas. We found no evidence of a strong founder effect in the new population; indeed genetic diversity in the newly founded population tended to be higher than in some of the source regions. Similarly, formation of the new population did not increase among-population genetic variance, and there was no evidence of kin-structured dispersal in the new population. These results indicate that high gene flow among sources and long-distance dispersal were important processes shaping the genetic diversity in this young V. membranaceum population. Other species with similar dispersal abilities may also be able to colonize new habitats without significant reduction in genetic diversity or increase in differentiation among populations. PMID:18194163
Genetic Engineering of Algae for Enhanced Biofuel Production ?
Radakovits, Randor; Jinkerson, Robert E.; Darzins, Al; Posewitz, Matthew C.
There are currently intensive global research efforts aimed at increasing and modifying the accumulation of lipids, alcohols, hydrocarbons, polysaccharides, and other energy storage compounds in photosynthetic organisms, yeast, and bacteria through genetic engineering. Many improvements have been realized, including increased lipid and carbohydrate production, improved H2 yields, and the diversion of central metabolic intermediates into fungible biofuels. Photosynthetic microorganisms are attracting considerable interest within these efforts due to their relatively high photosynthetic conversion efficiencies, diverse metabolic capabilities, superior growth rates, and ability to store or secrete energy-rich hydrocarbons. Relative to cyanobacteria, eukaryotic microalgae possess several unique metabolic attributes of relevance to biofuel production, including the accumulation of significant quantities of triacylglycerol; the synthesis of storage starch (amylopectin and amylose), which is similar to that found in higher plants; and the ability to efficiently couple photosynthetic electron transport to H2 production. Although the application of genetic engineering to improve energy production phenotypes in eukaryotic microalgae is in its infancy, significant advances in the development of genetic manipulation tools have recently been achieved with microalgal model systems and are being used to manipulate central carbon metabolism in these organisms. It is likely that many of these advances can be extended to industrially relevant organisms. This review is focused on potential avenues of genetic engineering that may be undertaken in order to improve microalgae as a biofuel platform for the production of biohydrogen, starch-derived alcohols, diesel fuel surrogates, and/or alkanes. PMID:20139239
Genetic engineering of phytochrome biosynthesis in bacteria
Gambetta, Gregory A.; Lagarias, J. Clark
The bilin prosthetic groups of the phytochrome photoreceptors and the light-harvesting phycobiliprotein antennae arise from the oxygen-dependent ring opening of heme. Two ferredoxin-dependent enzymes contribute to this conversion: a heme oxygenase and a bilin reductase with discrete double-bond specificity. Using a dual plasmid system, one expressing a truncated cyanobacterial apophytochrome 1, Cph1(N514), and the other expressing a two-gene operon consisting of a heme oxygenase and a bilin reductase, these studies establish the feasibility of producing photoactive phytochromes in any heme-containing cell. Heterologous expression systems for phytochromes not only will facilitate genetic analysis of their assembly, spectrophotometric activity, and biological function, but also might afford the means to regulate gene expression by light in nonplant cells. PMID:11553807
Gene Therapy in Large Animal Models of Human Cardiovascular Genetic Disease
Meg M. Sleeper; Lawrence T. Bish; H. Lee Sweeney
Several naturally occurring animal models for human genetic heart diseases offer an excellent opportunity to evaluate poten- tial novel therapies, including gene therapy. Some of these diseasesŚespecially those that result in a structural defect during development (e.g., patent ductus arteriosus, pulmonic stenosis)Śwould likely be diffi cult to treat with a therapeutic gene transfer approach. However, the ability to transduce a
Animal models of human genetic diseases: do they need to be faithful to be useful?
Jean-Louis Guķnet
With the advances in molecular genetics, animal models of human diseases are becoming more numerous and more refined every\\u000a year. Despite this, one must recognize that they generally do not faithfully and comprehensively mimic the homologous human\\u000a disease. Faced with these imperfections, some geneticists believe that these models are of little value, while for others,\\u000a on the contrary, they are
Genetic Evaluation of Dairy Goats for Milk and Fat Yield With an Animal Model1
G. R. Wiggans; J. W. J. Van Dijk; I. Misztal
An animal model was applied to evalu- ate 120,073 Alpine, LaMancha, Nubian, Saanen, and Toggenburg bucks and does. Parities higher than six were excluded. The model included fixed herd-year- season and random herd-sire interaction, permanent environmental breeding value, and residual effects. Breeding value in- cluded additive genetic value and fixed accumulated group effect. A doe's re- cords in different herds
Experimental investigations on the use of preheated animal fat as fuel in a compression ignition engine
M. Senthil Kumar; A. Kerihuel; J. Bellettre; M. Tazerout
The effect of fuel inlet temperature on performance, emission and combustion characteristics of a diesel engine is evaluated. A single cylinder direct injection diesel engine developing a power output of 2.8kW at 1500rev\\/min is tested using preheated animal fat as fuel. Experiments are conducted at the fuel inlet temperatures of 30, 40, 50, 60 and 70░C. Animal fat at low
A songbird animal model for dissecting the genetic bases of autism spectrum disorder.
Panaitof, S Carmen
The neural and genetic bases of human language development and associated neurodevelopmental disorders, including autism spectrum disorder (ASD), in which language impairment represents a core deficit, are poorly understood. Given that no single animal model can fully capture the behavioral and genetic complexity of ASD, work in songbird, an experimentally tractable animal model of vocal learning, can complement the valuable tool of rodent genetic models and contribute important insights to our understanding of the communication deficits observed in ASD. Like humans, but unlike traditional laboratory animals such as rodents or non-human primates, songbirds exhibit the capacity of vocal learning, a key subcomponent of language. Human speech and birdsong reveal important parallels, highlighting similar developmental critical periods, a homologous cortico-basal ganglia-thalamic circuitry, and a critical role for social influences in the learning of vocalizations. Here I highlight recent advances in using the songbird model to probe the cellular and molecular mechanisms underlying the formation and function of neural circuitry for birdsong and, by analogy, human language, with the ultimate goal of identifying any shared or human unique biological pathways underscoring language development and its disruption in ASD. PMID:22960335
A tissue engineering approach to bone repair in large animal models and in clinical practice
Ranieri Cancedda; Paolo Giannoni; Maddalena Mastrogiacomo
The repair of large segmental bone defects due to trauma, inflammation and tumor surgery remains a major clinical problem. Animal models were developed to test bone repair by tissue engineering approaches, mimicking real clinical situations. Studies differed with regard to animals (dog, sheep, goat), treated bone (femur, tibia, mandible), chemistry and structure of the scaffolds. Still, an advantage in the
Diverse plant and animal genetic records from Holocene and Pleistocene sediments.
Willerslev, Eske; Hansen, Anders J; Binladen, Jonas; Brand, Tina B; Gilbert, M Thomas P; Shapiro, Beth; Bunce, Michael; Wiuf, Carsten; Gilichinsky, David A; Cooper, Alan
Genetic analyses of permafrost and temperate sediments reveal that plant and animal DNA may be preserved for long periods, even in the absence of obvious macrofossils. In Siberia, five permafrost cores ranging from 400,000 to 10,000 years old contained at least 19 different plant taxa, including the oldest authenticated ancient DNA sequences known, and megafaunal sequences including mammoth, bison, and horse. The genetic data record a number of dramatic changes in the taxonomic diversity and composition of Beringian vegetation and fauna. Temperate cave sediments in New Zealand also yielded DNA sequences of extinct biota, including two species of ratite moa, and 29 plant taxa characteristic of the prehuman environment. Therefore, many sedimentary deposits may contain unique, and widespread, genetic records of paleoenvironments. PMID:12702808
Bilayer fibril formation by genetically engineered polypeptides: preparation and characterization.
Topilina, Natalya I; Higashiya, Seiichiro; Rana, Narender; Ermolenkov, Vladimir V; Kossow, Christopher; Carlsen, Autumn; Ngo, Silvana C; Wells, Christopher C; Eisenbraun, Eric T; Dunn, Kathleen A; Lednev, Igor K; Geer, Robert E; Kaloyeros, Alain E; Welch, John T
A de novo, genetically engineered 687 residue polypeptide expressed in E. coli has been found to form highly rectilinear, beta-sheet containing fibrillar structures. Tapping-mode atomic force microscopy, deep-UV Raman spectroscopy, and transmission electron microscopy definitively established the tendency of the fibrils to predominantly display an apparently planar bilayer or ribbon assemblage. The ordered self-assembly of designed, extremely repetitive, high molecular weight peptides is a harbinger of the utility of similar materials in nanoscience and engineering applications. PMID:16602727
Detecting Instability in Animal Social Networks: Genetic Fragmentation Is Associated with Social Instability in Rhesus Macaques
Beisner, Brianne A.; Jackson, Megan E.; Cameron, Ashley N.; McCowan, Brenda
The persistence of biological systems requires evolved mechanisms which promote stability. Cohesive primate social groups are one example of stable biological systems, which persist in spite of regular conflict. We suggest that genetic relatedness and its associated kinship structure are a potential source of stability in primate social groups as kinship structure is an important organizing principle in many animal societies. We investigated the effect of average genetic relatedness per matrilineal family on the stability of matrilineal grooming and agonistic interactions in 48 matrilines from seven captive groups of rhesus macaques. Matrilines with low average genetic relatedness show increased family-level instability such as: more sub-grouping in their matrilineal groom network, more frequent fighting with kin, and higher rates of wounding. Family-level instability in multiple matrilines within a group is further associated with group-level instability such as increased wounding. Stability appears to arise from the presence of clear matrilineal structure in the rhesus macaque group hierarchy, which is derived from cohesion among kin in their affiliative and agonistic interactions with each other. We conclude that genetic relatedness and kinship structure are an important source of group stability in animal societies, particularly when dominance and/or affilative interactions are typically governed by kinship. PMID:21298105
Role of gene therapy in tissue engineering procedures in rheumatology: the use of animal models
Peter M. van der Kraan; Wim B. van den Berg
Tissue engineering is not only the application of cells and scaffolds to generate a new tissue but should also bring into play biological principles to guide cellular behavior. A way to modify cellular behavior is genetic modification of the cells used for tissue engineering (gene therapy). In the field of rheumatic diseases, cellular modification by overexpressing anabolic factors, such as
Genetically engineered luminescent proteins in biosensing
NASA Astrophysics Data System (ADS)
Rowe, Laura; Ensor, Mark; Scott, Daniel; Deo, Sapna; Daunert, Sylvia
Luminescent proteins originally isolated from marine or terrestrial organisms have played a key role in the development of several biosensing systems. These proteins have been used in a variety of applications including, immunoassays, binding assays, cell-based sensing, high throughput screening, optical imaging, etc. Among the luminescent proteins isolated, the bioluminescent protein aequorin has been one of the proteins at the forefront in terms of its use in a vast number of biosensing systems. In our laboratory, we have employed aequorin as a label in the development of highly sensitive assays through chemical and genetic modifications from single step analysis of physiologically important molecules in biological fluids. An important aspect of optimizing these assays for clinical use involves understanding the stability of the various aequorin variants that are available. To this end we have designed several stability studies involving three important aequorin mutants, Mutant S, Mutant 5, and Mutant 53. The cysteine free aequorin, Mutant S, has been the most ubiquitously used aequorin variant in our laboratory because of its increased stability and activity as compared to native aequorin. Mutant 5 and Mutant 53 contain a single cyteine residue at position 5 and 53 in the protein, respectively. Because of the presence of a single cysteine residue, Mutant 5 and Mutant 53 both can be site-specifically conjugated. This site specific conjugation capability gives Mutant 5 and Mutant 53 an advantage over native aequorin when developing assays. Additional studies optimizing the expression, purification, and charging of aequorin Mutant S were also performed. A thorough understanding of the efficient expression, purification, and storage of these aequorin mutants will allow for the more practical utilization of these mutants in the development of future biosensing systems.
76 FR 78232 - Monsanto Co.; Determination of Nonregulated Status for Soybean Genetically Engineered To Have a...
Federal Register 2010, 2011, 2012, 2013, 2014
...OF AGRICULTURE Animal and Plant Health Inspection Service...Glyphosate AGENCY: Animal and Plant Health Inspection Service...soybeans are likely to pose a plant pest risk, the draft EA...tests, and health effects of genetically modified organisms and...
What are the prospects for genetically engineered, disease resistant plants?
David B. Collinge; Ole S°gaard Lund; Hans Thordal-Christensen
Insect and herbicide-resistant plants are the most widely grown transgenics in agricultural production. No strategy using\\u000a genetically engineered plants for disease resistance has had a comparable impact. Why is this? What are the prospects for\\u000a introducing transgenic disease resistant plants to agriculture? We review the biological background for strategies used to\\u000a make disease resistant GM crops, illustrate examples of these
Oncolytic virus therapy using genetically engineered herpes simplex viruses
Tomoki Todo
An increasing number of oncolytic virus vectors has been developed lately for cancer therapy. Herpes simplex virus type 1\\u000a (HSV-1) vectors are particularly useful, because they can be genetically engineered to replicate and spread highly selectively\\u000a in tumor cells and can also express multiple foreign transgenes. These vectors can manifest cytopathic effect in a wide variety\\u000a of tumor types without
Prevention of preharvest aflatoxin contamination through genetic engineering of crops
K. Rajasekaran; J. W. Cary; T. E. Cleveland
Current practices on prevention of aflatoxin contamination of crop species include time consuming, expensive agronomic practices.\\u000a Of all the methods available to-date, conventional breeding and\\/or genetic engineering to develop host plant-based resistance\\u000a to aflatoxin-producing fungi appear to be valuable for several reasons. However, breeding for disease-resistant crops is very\\u000a time consuming, especially in tree crops, and does not lend itself
Utilization of farm animal genetic resources in a changing agro-ecological environment in the Nordic countries
Kantanen, Juha; L°vendahl, Peter; Strandberg, Erling; Eythorsdottir, Emma; Li, Meng-Hua; Kettunen-Prµbel, Anne; Berg, Peer; Meuwissen, Theo
Livestock production is the most important component of northern European agriculture and contributes to and will be affected by climate change. Nevertheless, the role of farm animal genetic resources in the adaptation to new agro-ecological conditions and mitigation of animal productionÆs effects on climate change has been inadequately discussed despite there being several important associations between animal genetic resources and climate change issues. The sustainability of animal production systems and future food security require access to a wide diversity of animal genetic resources. There are several genetic questions that should be considered in strategies promoting adaptation to climate change and mitigation of environmental effects of livestock production. For example, it may become important to choose among breeds and even among farm animal species according to their suitability to a future with altered production systems. Some animals with useful phenotypes and genotypes may be more useful than others in the changing environment. Robust animal breeds with the potential to adapt to new agro-ecological conditions and tolerate new diseases will be needed. The key issue in mitigation of harmful greenhouse gas effects induced by livestock production is the reduction of methane (CH4) emissions from ruminants. There are differences in CH4 emissions among breeds and among individual animals within breeds that suggest a potential for improvement in the trait through genetic selection. Characterization of breeds and individuals with modern genomic tools should be applied to identify breeds that have genetically adapted to marginal conditions and to get critical information for breeding and conservation programs for farm animal genetic resources. We conclude that phenotyping and genomic technologies and adoption of new breeding approaches, such as genomic selection introgression, will promote breeding for useful characters in livestock species. PMID:25767477
In order to build up a system of international cooperative research in the field of animal biotechnology, animal stem cell engineering and healthcare biotechnology, the Graduate School
Takahashi, Ryo
biotechnology, animal stem cell engineering and healthcare biotechnology, the Graduate School of Bioagricultural Spin-offs in Animal Biotechnology Project of Educational and Research Collaborative Internship Program in Animal Biotechnology Graduate School of Bioagricultural Sciences, Nagoya University Furo-cho, Chikusa
Versatile RNA-sensing transcriptional regulators for engineering genetic networks
Lucks, Julius B.; Qi, Lei; Mutalik, Vivek K.; Wang, Denise; Arkin, Adam P.
The widespread natural ability of RNA to sense small molecules and regulate genes has become an important tool for synthetic biology in applications as diverse as environmental sensing and metabolic engineering. Previous work in RNA synthetic biology has engineered RNA mechanisms that independently regulate multiple targets and integrate regulatory signals. However, intracellular regulatory networks built with these systems have required proteins to propagate regulatory signals. In this work, we remove this requirement and expand the RNA synthetic biology toolkit by engineering three unique features of the plasmid pT181 antisense-RNA-mediated transcription attenuation mechanism. First, because the antisense RNA mechanism relies on RNA-RNA interactions, we show how the specificity of the natural system can be engineered to create variants that independently regulate multiple targets in the same cell. Second, because the pT181 mechanism controls transcription, we show how independently acting variants can be configured in tandem to integrate regulatory signals and perform genetic logic. Finally, because both the input and output of the attenuator is RNA, we show how these variants can be configured to directly propagate RNA regulatory signals by constructing an RNA-meditated transcriptional cascade. The combination of these three features within a single RNA-based regulatory mechanism has the potential to simplify the design and construction of genetic networks by directly propagating signals as RNA molecules. PMID:21555549
Genetic engineering: a matter that requires further refinement in Spanish secondary school textbooks
M. V. MartĒnez-Gracia; M. J. Gil-Qu²lez; J. Osada
Genetic engineering is now an integral part of many high school textbooks but little work has been done to assess whether it is being properly addressed. A checklist with 19 items was used to analyze how genetic engineering is presented in biology textbooks commonly used in Spanish high schools, including the content, its relationship with fundamental genetic principles, and how
Genetic improvement of reproduction : time for deeds A.R.C. Animal Breeding Research Organisation, West Mains Road,
Paris-Sud XI, Universitķ de
Genetic improvement of reproduction : time for deeds R.B. LAND A.R.C. Animal Breeding Research by genetic selection within as well as among populations. The availability of diverse lines would facilitate selection among populations and serve as the basis for research to assess criteria to increase the accuracy
Optimisation of the Gas-Exchange System of Combustion Engines by Genetic Algorithm
Marsland, Stephen
Optimisation of the Gas-Exchange System of Combustion Engines by Genetic Algorithm C. D. Rose, S. R of combustion engine gas-exchange systems still predominantly use trial and error. This paper proposes a new. Keywords - genetic algorithm; variable-length input encoding; combustion engine; optimisation I
Research review paper Genome-scale genetic engineering in Escherichia coli
Bang, Duhee
Research review paper Genome-scale genetic engineering in Escherichia coli Jaehwan Jeong 1 , Namjin cells. Escherichia coli has been a particularly good model organism for bac- terial genome engineering Keywords: Genome engineering Red recombination Multiplex automated genome engineering Genome engineering
ERIC Educational Resources Information Center
Tech Directions, 2008
Art and animation work is the most significant part of electronic game development, but is also found in television commercials, computer programs, the Internet, comic books, and in just about every visual media imaginable. It is the part of the project that makes an abstract design idea concrete and visible. Animators create the motion of life inģ
Contribution of genetic influences to animal-to-animal variation in myoglobin content and beef lean color stability.
King, D A; Shackelford, S D; Kuehn, L A; Kemp, C M; Rodriguez, A B; Thallman, R M; Wheeler, T L
Longissimus thoracis steaks from steers (n = 464) with 0 to 50% inheritance of Angus, Charolais, Gelbvieh, Hereford, Limousin, Red Angus, and Simmental were evaluated during 6 d of display to assess genetic contributions to color stability. Color space values [CIE L* (lightness), a* (redness), b* (yellowness)], chroma, color change (DeltaE), and surface metmyoglobin (K/S 572/525) were determined on d 0 and 6 of display. Myoglobin concentration was highly heritable (0.85), but ultimate pH was weakly heritable (0.06). Day 0 L* values were moderately heritable (0.24). Variation in metmyoglobin, L*, and DeltaE on d 6 was moderately explained by genetic factors (41, 40, and 29%, respectively). Change during display was moderately heritable for a* (0.31), b* (0.23), chroma (0.35), and surface metmyoglobin (0.29). At the start of display, Angus steaks had greater (P < 0.05) L* values than those from all breeds except Charolais. On d 6, Angus steaks had greater (P < 0.05) L* (50.0) values than Gelbvieh, Hereford, and Simmental steaks (46.1, 44.0, and 44.5, respectively). Day 0 values for a*, b*, chroma, and DeltaE were not affected by breed (P > 0.05). On d 6, a* values were greater (P < 0.05) for Charolais and Limousin steaks (31.1 and 30.5) than Angus, Hereford, and Red Angus steaks (27.4, 27.7, and 26.3, respectively). Thus, a* changed less (P < 0.05) in Charolais and Limousin steaks (1.8 and 2.6, respectively) vs. steaks from other breeds. Day 6 b* values were greater (P < 0.05) in Charolais (24.5) and Limousin steaks (24.0) vs. Gelbvieh (22.2), Hereford (21.9), and Red Angus steaks (21.4). Thus, b* values changed less (P < 0.05) in Charolais and Limousin steaks (1.5 and 1.7, respectively) than in Angus, Gelbvieh, Hereford, and Red Angus steaks (4.3, 3.8, 4.4, and 5.1, respectively). After 6 d of display, Charolais and Limousin steaks had greater chroma (P < 0.05; 39.5 and 38.8, respectively) compared with Angus, Hereford, and Red Angus steaks (35.4, 35.3, and 33.9, respectively). Less (P < 0.05) change in chroma occurred for Charolais and Limousin (2.1 and 2.8, respectively) than in Angus, Gelbvieh, Hereford, and Red Angus steaks (7.1, 6.6, 7.4, and 9.0, respectively). Myoglobin concentration was less for Charolais and Limousin (P < 0.05; 2.77 and 2.72, respectively) compared with Gelbvieh, Red Angus, and Simmental steaks (3.62, 3.43, and 3.71, respectively). Breeds did not differ in pH (P > 0.05). These data suggest Charolais- and Limousin-carcasses produced steaks with greater lean color stability than Angus, Hereford, and Red Angus carcasses. Furthermore, these findings suggest that genetics contribute substantially to animal-to-animal variation in lean color, particularly in maintaining color. PMID:19966159
476. Human Mesenchymal Stem Cells Genetically Modified To Express High Levels of Erythropoietin Through Transfer of an Artificial Chromosome Engineered Using the ACE System
Sandra Vanderbyl; Neil MacDonald; Tom Stodola; Adele Telenius; Sandra Stewart; Harry C. Ledebur; Carl F. Perez
The ACE System is a versatile, reliable system for genetically modifying cell therapies, generating transgenic animals, and engineering mammalian cells for high expression of a recombinant protein. Key components of the ACE System include an artificial chromosome (Platform ACE) encoding multiple (>50) DNA site-specific integration sites (acceptor sites); a targeting vector (ACE Targeting Vector) encoding both a Platform ACE-specific DNA
Recombineering: genetic engineering in bacteria using homologous recombination.
Thomason, Lynn; Court, Donald L; Bubunenko, Mikail; Costantino, Nina; Wilson, Helen; Datta, Simanti; Oppenheim, Amos
The bacterial chromosome and plasmids can be engineered in vivo by homologous recombination using PCR products and synthetic oligonucleotides as substrates. This is possible because bacteriophage-encoded recombination functions efficiently to recombine sequences with homologies as short as 35 to 40 bases. This recombineering allows DNA sequences to be inserted or deleted without regard to location of restriction sites. This unit first describes preparation of electrocompetent cells expressing the recombineering functions and their transformation with dsDNA or ssDNA. Support protocols describe a two-step method of making genetic alterations without leaving any unwanted changes, and a method for retrieving a genetic marker (cloning) from the E. coli chromosome or a co-electroporated DNA fragment and moving it onto a plasmid. A method is also given to screen for unselected mutations. Additional protocols describe removal of defective prophage, methods for recombineering. PMID:18265390
Multi-engine Animal Disease Diagnostic Expert System
TAN Wen-xue; GUO Guo-qiang; WANG Jing-ren
?Abstract?The traditional disease diagnostic expertsy stemonly has a single reasoning process to use knowledge,so that its knowledge utilization is inefficient. Its conclusion has low accuracyand without contrast. This paper makes example ofgoat, modelings disease diagnostic knowledge base by the use of object-oriented knowledge representation. According to the training process and diagnostic model, it proposes the ideology of multi-engine ruling based
Stereo Visualized Animation Content for the Learning of the Mechanism of Engine
NASA Astrophysics Data System (ADS)
Sato, Tomoaki
The stereo visualized animation contents have not been spread on the domain of education because of the difficulties and high cost of making that. On this study, we proposed an easy and low cost method of the stereo visualization for 3DCG-Animation. The easy and low cost stereo visualization system is composed of two ordinary liquid crystal projectors, a personal computer and a silver screen. In this paper, by using this method, we developed a stereo visualized 3DCG-animation content of 4 stroke cycle gasoline engine and used the content for the class of the machining practice exercise. Moreover, the learning effect of the content was examined with the questionnaire. The result of questionnaire showed that the stereo visualized 3DCG animation content was very helpful to understand the mechanism of engine.
Genetic engineering: Rifkin strikes at corn this time.
Budiansky, S
As a result of a threatened suit by Jeremy Rifkin, Stanford University has postponed an experiment involving a test plot of genetically-engineered corn. At issue is an injunction forbidding the Recombinant DNA Advisory Committee of the National Institutes of Health from approving federal funding of experiments entailing the release of recombinant DNA into the environment. Rifkin's legal argument is that an environmnental impact statement must be filed for both commercially- and federally-funded research. It is expected that Rifkin's demand for equal treatment regardless of funding source will be agreed to by NIH. PMID:6588294
Hybrid anthracyclines from a genetically engineered Streptomyces galilaeus mutant.
Kunnari, T J; Ylihonko, K P; Klika, K D; Mõntsõlõ, P I; Hakala, J M
The genetic engineering of antibiotic-producing Streptomyces strains is an approach that is emerging and ready to become established as a successful methodology in developing analogues of the original, pharmaceutically important, natural products obtained from the organisms. The current report highlights this succes by demonstrating the high-level production of novel anthracyclines. The biosynthetic pathways of the nogalamycin-producing Streptomyces nogalater and the aclacinomycin-producing S. galilaeus were combined by transferring the genes of S. nogalater polyketide synthetase into a nonproducing S. galilaeus mutant. The resulting anthracycline antibiotics that were produced possessed structural features characteristic of compounds from both of the undoctored Streptomycesstrains. PMID:10814169
A 3D character animation engine for multimodal interaction on mobile devices
Enrico Sandali; Fabio Lavagetto; Paolo Pisano
Talking virtual characters are graphical simulations of real or imaginary persons that enable natural and pleasant multimodal interaction with the user, by means of voice, eye gaze, facial expression and gestures. This paper presents an implementation of a 3D virtual character animation and rendering engine, compliant with the MPEG-4 standard, running on Symbian-based SmartPhones. Real-time animation of virtual characters on
Challenges to IPM Advancement: Pesticides, Biocontrol, Genetic Engineering, and Invasive Species.
Hoddle, Mark S.
77 Challenges to IPM Advancement: Pesticides, Biocontrol, Genetic Engineering, and Invasive Species of transgenic crops, invasive species, and a diminishing base of scientific talent and research funding become
NSDL National Science Digital Library
Christopher Griffith
This collection contains animations of a nuclear chain reaction, nuclear fission and nuclear fusion. It also showcases interactive models of the first atomic bombs and simulation of the "Nuclear Winter" effect.
Cancer Regression in Patients After Transfer of Genetically Engineered Lymphocytes
NASA Astrophysics Data System (ADS)
Morgan, Richard A.; Dudley, Mark E.; Wunderlich, John R.; Hughes, Marybeth S.; Yang, James C.; Sherry, Richard M.; Royal, Richard E.; Topalian, Suzanne L.; Kammula, Udai S.; Restifo, Nicholas P.; Zheng, Zhili; Nahvi, Azam; de Vries, Christiaan R.; Rogers-Freezer, Linda J.; Mavroukakis, Sharon A.; Rosenberg, Steven A.
Through the adoptive transfer of lymphocytes after host immunodepletion, it is possible to mediate objective cancer regression in human patients with metastatic melanoma. However, the generation of tumor-specific T cells in this mode of immunotherapy is often limiting. Here we report the ability to specifically confer tumor recognition by autologous lymphocytes from peripheral blood by using a retrovirus that encodes a T cell receptor. Adoptive transfer of these transduced cells in 15 patients resulted in durable engraftment at levels exceeding 10% of peripheral blood lymphocytes for at least 2 months after the infusion. We observed high sustained levels of circulating, engineered cells at 1 year after infusion in two patients who both demonstrated objective regression of metastatic melanoma lesions. This study suggests the therapeutic potential of genetically engineered cells for the biologic therapy of cancer.
Genetic aspects of risk - species diversification, genetic management and genetic engineering
Rowland D. Burdon
The topic is addressed in terms of (1) risk profiles (2) the types of risk management measure taken: risk avoidance, risk spread, and response preparation (3) types of risk to be addressed: biological and market (4) the levels in a genetic hierarchy at which measŁ ures are taken: species, provenances, breeds, indiŁ vidual genotypes, and individual genes. Within this framework,
Tissue Engineering in Animal Models for Urinary Diversion: A Systematic Review
Sloff, Marije; de Vries, Rob; Geutjes, Paul; IntHout, Joanna; Ritskes-Hoitinga, Merel
Tissue engineering and regenerative medicine (TERM) approaches may provide alternatives for gastrointestinal tissue in urinary diversion. To continue to clinically translatable studies, TERM alternatives need to be evaluated in (large) controlled and standardized animal studies. Here, we investigated all evidence for the efficacy of tissue engineered constructs in animal models for urinary diversion. Studies investigating this subject were identified through a systematic search of three different databases (PubMed, Embase and Web of Science). From each study, animal characteristics, study characteristics and experimental outcomes for meta-analyses were tabulated. Furthermore, the reporting of items vital for study replication was assessed. The retrieved studies (8 in total) showed extreme heterogeneity in study design, including animal models, biomaterials and type of urinary diversion. All studies were feasibility studies, indicating the novelty of this field. None of the studies included appropriate control groups, i.e. a comparison with the classical treatment using GI tissue. The meta-analysis showed a trend towards successful experimentation in larger animals although no specific animal species could be identified as the most suitable model. Larger animals appear to allow a better translation to the human situation, with respect to anatomy and surgical approaches. It was unclear whether the use of cells benefits the formation of a neo urinary conduit. The reporting of the methodology and data according to standardized guidelines was insufficient and should be improved to increase the value of such publications. In conclusion, animal models in the field of TERM for urinary diversion have probably been chosen for reasons other than their predictive value. Controlled and comparative long term animal studies, with adequate methodological reporting are needed to proceed to clinical translatable studies. This will aid in good quality research with the reduction in the use of animals and an increase in empirical evidence of biomedical research. PMID:24964011
Genetic effects of contaminant exposure--towards an assessment of impacts on animal populations.
Hebert, P D; Luiker, M M
This review aims both to identify the potential risks to animal populations as a consequence of exposure to genotoxins and to identify the techniques most useful in assessing these risks. These evaluations are complicated by the fact that contaminant exposure acts both to restructure naturally occurring genetic diversity and, when contaminants have mutagenic activity, to enhance the rate of introduction of new variation. There is now evidence that contaminant exposure often leads to change in the genetic attributes of natural populations. Short-lived organisms often develop resistance to contaminants, with only modest impacts on diversity in the balance of the genome, although massive mortality occurs during the gene replacement. Resistance is, however, less likely to evolve in species with small population size, such as many wildlife species. Such species will experience population declines or extinction as the impact of contaminants on physiological systems is not counteracted by gene replacements. Even when adaptation to exposure occurs, populations may suffer diminished fitness as a consequence of the mutagenic effects of contaminants. The expression of these effects range from an increase in the incidence of developmental abnormalities to shifts in chromosomal and gene structure. The assessment of this broad range of impacts can only be accomplished with a spectrum of analytical approaches. However, recent advances in molecular and developmental genetics are now making possible the detailed assessment of these mutagenic impacts in natural populations. PMID:8885423
A Knockout Experiment: Disciplinary Divides and Experimental Skill in Animal Behaviour Genetics.
Whitfield, Nicholas; Schlich, Thomas
In the early 1990s, a set of new techniques for manipulating mouse DNA allowed researchers to 'knock out' specific genes and observe the effects of removing them on a live mouse. In animal behaviour genetics, questions about how to deploy these techniques to study the molecular basis of behaviour became quite controversial, with a number of key methodological issues dissecting the interdisciplinary research field along disciplinary lines. This paper examines debates that took place during the 1990s between a predominately North American group of molecular biologists and animal behaviourists around how to design, conduct, and interpret behavioural knockout experiments. Drawing from and extending Harry Collins's work on how research communities negotiate what counts as a 'well-done experiment,' I argue that the positions practitioners took on questions of experimental skill reflected not only the experimental traditions they were trained in but also their differing ontological and epistemological commitments. Different assumptions about the nature of gene action, eg., were tied to different positions in the knockout mouse debates on how to implement experimental controls. I conclude by showing that examining representations of skill in the context of a community's knowledge commitments sheds light on some of the contradictory ways in which contemporary animal behaviour geneticists talk about their own laboratory work as a highly skilled endeavour that also could be mechanised, as easy to perform and yet difficult to perform well. PMID:26090739
Priceless GEMMs: genetically engineered mouse models for colorectal cancer drug development.
Roper, Jatin; Hung, Kenneth E
To establish effective drug development for colorectal cancer (CRC), preclinical models that are robust surrogates for human disease are crucial. Mouse models are an attractive platform because of their relatively low cost, short life span, and ease of use. There are two main categories of mouse CRC models: xenografts derived from implantation of CRC cells or tumors in immunodeficient mice; and genetically engineered mouse models (GEMMs) derived from modification of human cancer predisposition genes, resulting in spontaneous tumor formation. Here, we review xenografts and GEMMs and focus on their potential application in translational research. Furthermore, we describe newer GEMMs for sporadic CRC that are particularly suitable for drug testing. Finally, we discuss recent advances in small-animal imaging, such as optical colonoscopy, which allow in vivo assessment of tumors. With the increasing sophistication of GEMMs, our preclinical armamentarium provides new hope for the ongoing war against CRC. PMID:22739258
Teaching Habitat and Animal Classification to Fourth Graders Using an Engineering-Design Model
ERIC Educational Resources Information Center
Marulcu, Ismail
Background: The motivation for this work is built upon the premise that there is a need for research-based materials for design-based science instruction. In this paper, a small portion of our work investigating the impact of a LEGO[TM] engineering unit on fourth grade students' preconceptions and understanding of animals is presented.ģ
Genetic Engineering: A Matter that Requires Further Refinement in Spanish Secondary School Textbooks
ERIC Educational Resources Information Center
Martinez-Gracia, M. V.; Gil-Quylez, M. J.; Osada, J.
Genetic engineering is now an integral part of many high school textbooks but little work has been done to assess whether it is being properly addressed. A checklist with 19 items was used to analyze how genetic engineering is presented in biology textbooks commonly used in Spanish high schools, including the content, its relationship withģ
Gender Differences in the Perception of Genetic Engineering Applied to Human Reproduction
Carol L. Napolitano; Oladele A. Ogunseitan
A questionnaire-survey of public perception of the desirability, risks, and benefits associated with current and potential applications of genetic engineering techniques to manipulate the outcome of human reproduction was conducted on 111 male and 135 female respondents. The proportion (63%) of male respondents who hold a positive impression that genetic engineering is a socially beneficial field of scientific research was
Dynamics of Horizontal Gene Transfer and the Ecological Risk Assessment of Genetically Engineered Organisms
Wayne G. Landis; Lori A. Lenart; Julann A. Spromberg
The extensive published discussion of potential ecological impacts of introduced genetic sequences and genetically engineered organisms has lacked a quantified delineation of the critical questions for the estimation of risk. Ultimately, the ecological risk assessment of introduced gene sequences is the application of evolution, population genetics, and ecology to risk estimation and decision making. This paper provides a framework for
76 FR 63278 - Bayer CropScience LP; Determination of Nonregulated Status for Cotton Genetically Engineered for...
Federal Register 2010, 2011, 2012, 2013, 2014
...Determination of Nonregulated Status for Cotton Genetically Engineered for Insect Resistance...determination that a genetically engineered cotton developed by Bayer CropScience LP, designated as TwinLink TM cotton (events T304-40 and GHB119),...
The agricultural use of genetically engineered plants and microorganisms has become increasingly common. Because genetically engineered plants and microorganisms can produce compounds foreign to their environment, there is concern that they may become established outside of thei...
Recombineering: a homologous recombination-based method of genetic engineering.
Sharan, Shyam K; Thomason, Lynn C; Kuznetsov, Sergey G; Court, Donald L
Recombineering is an efficient method of in vivo genetic engineering applicable to chromosomal as well as episomal replicons in Escherichia coli. This method circumvents the need for most standard in vitro cloning techniques. Recombineering allows construction of DNA molecules with precise junctions without constraints being imposed by restriction enzyme site location. Bacteriophage homologous recombination proteins catalyze these recombineering reactions using double- and single-stranded linear DNA substrates, so-called targeting constructs, introduced by electroporation. Gene knockouts, deletions and point mutations are readily made, gene tags can be inserted and regions of bacterial artificial chromosomes or the E. coli genome can be subcloned by gene retrieval using recombineering. Most of these constructs can be made within about 1 week's time. PMID:19180090
Recombineering: A Homologous Recombination-Based Method of Genetic Engineering
Sharan, Shyam K.; Thomason, Lynn C.; Kuznetsov, Sergey G.; Court, Donald L.
Recombineering is an efficient method of in vivo genetic engineering applicable to chromosomal as well as episomal replicons in E. coli. This method circumvents the need for most standard in vitro cloning techniques. Recombineering allows construction of DNA molecules with precise junctions without constraints being imposed by restriction enzyme site location. Bacteriophage homologous recombination proteins catalyze these recombineering reactions using double- and single-strand linear DNA substrates, so-called targeting constructs, introduced by electroporation. Gene knockouts, deletions and point mutations are readily made, gene tags can be inserted, and regions of bacterial artificial chromosomes (BACs) or the E. coli genome can be subcloned by gene retrieval using recombineering. Most of these constructs can be made within about a week's time. PMID:19180090
Surveys suck: Consumer preferences when purchasing genetically engineered foods.
Powell, Douglas A
Many studies have attempted to gauge consumers' acceptance of genetically engineered or modified (GM) foods. Surveys, asking people about attitudes and intentions, are easy-to-collect proxies of consumer behavior. However, participants tend to respond as citizens of society, not discrete individuals, thereby inaccurately portraying their potential behavior. The Theory of Planned Behavior improved the accuracy of self-reported information, but its limited capacity to account for intention variance has been attributed to the hypothetical scenarios to which survey participants must respond. Valuation methods, asking how much consumers may be willing to pay or accept for GM foods, have revealed that consumers are usually willing to accept them at some price, or in some cases willing to pay a premium. Ultimately, it's consumers' actual--not intended--behavior that is of most interest to policy makers and business decision-makers. Real choice experiments offer the best avenue for revealing consumers' food choices in normal life. PMID:24281042
Genetic causes of transitions from sexual reproduction to asexuality in plants and animals.
Neiman, M; Sharbel, T F; Schwander, T
The persistence of sexual reproduction in the face of competition from asexual invaders is more likely if asexual lineages are produced infrequently or have low fitness. The generation rate and success of new asexual lineages will be influenced by the proximate mechanisms underlying transitions to asexuality. As such, characterization of these mechanisms can help explain the distribution of reproductive modes among natural populations. Here, we synthesize the literature addressing proximate causes of transitions from sexual to asexual reproduction in plants and animals. In cyclical and facultatively asexual taxa, individual mutations can cause obligate asexuality. The evolution of asexuality in obligately sexual groups is more complex, requiring the simultaneous acquisition of two traits generally controlled by different genetic factors: unreduced gamete formation and spontaneous development of unfertilized gametes. At least three 'pre-adaptations' could favour transitions to obligate asexuality in obligate sexuals. First, linkage among loci affecting separate key components of asexuality facilitates its spread, with evidence for these linkage blocks in plants. Second, asexuality should evolve more readily in haplodiploids; support for this hypothesis comes from two examples where a single locus causes transitions to asexuality. Third, standing genetic variation for the production of unreduced gametes could facilitate transitions to asexuality, but whether the ability to produce unreduced gametes contributes to the evolution of obligate asexuality remains unclear. We close by reviewing the associations between asexuality, hybridization and polyploidy, and argue that current data suggest that hybridization is more likely to play a causal role in transitions to asexuality than polyploidy. PMID:24666600
Reward circuitry dysfunction in psychiatric and neurodevelopmental disorders and genetic syndromes: animal models and clinical findings
This review summarizes evidence of dysregulated reward circuitry function in a range of neurodevelopmental and psychiatric disorders and genetic syndromes. First, the contribution of identifying a core mechanistic process across disparate disorders to disease classification is discussed, followed by a review of the neurobiology of reward circuitry. We next consider preclinical animal models and clinical evidence of reward-pathway dysfunction in a range of disorders, including psychiatric disorders (i.e., substance-use disorders, affective disorders, eating disorders, and obsessive compulsive disorders), neurodevelopmental disorders (i.e., schizophrenia, attention-deficit/hyperactivity disorder, autism spectrum disorders, TouretteÆs syndrome, conduct disorder/oppositional defiant disorder), and genetic syndromes (i.e., Fragile X syndrome, Prader¢Willi syndrome, Williams syndrome, Angelman syndrome, and Rett syndrome). We also provide brief overviews of effective psychopharmacologic agents that have an effect on the dopamine system in these disorders. This review concludes with methodological considerations for future research designed to more clearly probe reward-circuitry dysfunction, with the ultimate goal of improved intervention strategies. PMID:22958744
Recombineering: genetic engineering in bacteria using homologous recombination.
Thomason, Lynn C; Sawitzke, James A; Li, Xintian; Costantino, Nina; Court, Donald L
The bacterial chromosome and bacterial plasmids can be engineered in vivo by homologous recombination using PCR products and synthetic oligonucleotides as substrates. This is possible because bacteriophage-encoded recombination proteins efficiently recombine sequences with homologies as short as 35 to 50 bases. Recombineering allows DNA sequences to be inserted or deleted without regard to location of restriction sites. This unit first describes preparation of electrocompetent cells expressing the recombineering functions and their transformation with dsDNA or ssDNA. It then presents support protocols that describe several two-step selection/counter-selection methods of making genetic alterations without leaving any unwanted changes in the targeted DNA, and a method for retrieving onto a plasmid a genetic marker (cloning by retrieval) from the Escherichia coli chromosome or a co-electroporated DNA fragment. Additional protocols describe methods to screen for unselected mutations, removal of the defective prophage from recombineering strains, and other useful techniques. Curr. Protoc. Mol. Biol. 106:1.16.1-1.16.39. ® 2014 by John Wiley & Sons, Inc. PMID:24733238
Novel method for monitoring genetically engineered microorganisms in the environment.
Chaudhry, G R; Toranzos, G A; Bhatti, A R
A method has been devised for directly detecting and monitoring genetically engineered microorganisms (GEMs) by using in vitro amplification of the target DNAs by a polymerase chain reaction and then hybridizing the DNAs with a specific oligonucleotide or DNA probe. A cloned 0.3-kilobase napier grass (Pennisetum purpureum) genomic DNA that did not hybridize to DNAs isolated from various microorganisms, soil sediments, and aquatic environments was inserted into a derivative of a 2,4-dichlorophenoxyacetic acid-degradative plasmid, pRC10, and transferred into Escherichia coli. This genetically altered microorganism, seeded into filter-sterilized lake and sewage water samples (10(4)/ml), was detected by a plate count method in decreasing numbers for 6 and 10 days of sample incubation, respectively. The new method detected the amplified unique marker (0.3-kilobase DNA) of the GEM even after 10 to 14 days of incubation. This method is highly sensitive (it requires only picogram amounts of DNA) and has an advantage over the plate count technique, which can detect only culturable microorganisms. The method may be useful for monitoring GEMs in complex environments, where discrimination between GEMs and indigenous microorganisms is either difficult or requires time-consuming tests. PMID:2667463
Exploring Dynamics of Molybdate in Living Animal Cells by a Genetically Encoded FRET Nanosensor
Nakanishi, Yoichi; Iida, Syuntaro; Ueoka-Nakanishi, Hanayo; Niimi, Tomoaki; Tomioka, Rie; Maeshima, Masayoshi
Molybdenum (Mo) is an essential trace element for almost all living organisms including animals. Mo is used as a catalytic center of molybdo-enzymes for oxidation/reduction reactions of carbon, nitrogen, and sulfur metabolism. Whilst living cells are known to import inorganic molybdate oxyanion from the surrounding environment, the in vivo dynamics of cytosolic molybdate remain poorly understood as no appropriate indicator is available for this trace anion. We here describe a genetically encoded F÷rester-resonance-energy-transfer (FRET)-based nanosensor composed of CFP, YFP and the bacterial molybdate-sensor protein ModE. The nanosensor MolyProbe containing an optimized peptide-linker responded to nanomolar-range molybdate selectively, and increased YFP:CFP fluorescence intensity ratio by up to 109%. By introduction of the nanosensor, we have been able to successfully demonstrate the real-time dynamics of molybdate in living animal cells. Furthermore, time course analyses of the dynamics suggest that novel oxalate-sensitive- and sulfate-resistant- transporter(s) uptake molybdate in a model culture cell. PMID:23472155
Genetically encoded molecular biosensors to image histone methylation in living animals.
Sekar, Thillai V; Foygel, Kira; Gelovani, Juri G; Paulmurugan, Ramasamy
Post-translational addition of methyl groups to the amino terminal tails of histone proteins regulates cellular gene expression at various stages of development and the pathogenesis of cellular diseases, including cancer. Several enzymes that modulate these post-translational modifications of histones are promising targets for development of small molecule drugs. However, there is no promising real-time histone methylation detection tool currently available to screen and validate potential small molecule histone methylation modulators in small animal models. With this in mind, we developed genetically encoded molecular biosensors based on the split-enzyme complementation approach for in vitro and in vivo imaging of lysine 9 (H3-K9 sensor) and lysine 27 (H3-K27 sensor) methylation marks of histone 3. These methylation sensors were validated in vitro in HEK293T, HepG2, and HeLa cells. The efficiency of the histone methylation sensor was assessed by employing methyltransferase inhibitors (Bix01294 and UNC0638), demethylase inhibitor (JIB-04), and siRNA silencing at the endogenous histone K9-methyltransferase enzyme level. Furthermore, noninvasive bioluminescence imaging of histone methylation sensors confirmed the potential of these sensors in monitoring histone methylation status in response to histone methyltransferase inhibitors in living animals. Experimental results confirmed that the developed H3-K9 and H3-K27 sensors are specific and sensitive to image the drug-induced histone methylation changes in living animals. These novel histone methylation sensors can facilitate the in vitro screening and in vivo characterization of new histone methyltransferase inhibitors and accelerate the pace of introduction of epigenetic therapies into the clinic. PMID:25506787
Anim. Behav., 1998, 55, 417426 Genetic effects on task performance, but not on age polyethism, in a
O'Donnell, Sean
Anim. Behav., 1998, 55, 417Ł426 Genetic effects on task performance, but not on age polyethism, in a swarm-founding eusocial wasp SEAN O'DONNELL Department of Psychology, University of Washington (Received-related changes in behaviour (age polyethism) and specialization in task performance. The aim of this study
Integrating policies for the management of animal genetic resources with demand for livestock products and environmental sustainability
Technology Transfer Automated Retrieval System (TEKTRAN)
Recognition of the need to conserve animal genetic resources comes at a time when the global livestock sector faces significant challenges in meeting the growing demand for livestock products and the mitigation of negative environmental impacts caused by livestock. Outside of the U.S. it would seem ...
Fear induced neuronal alterations in a genetic model of depression: An fMRI study on awake animals
Wei Huang; Meghan E. Heffernan; Zhixin Li; Nanyin Zhang; David H. Overstreet; Jean A. King
Previous human imaging studies used facial stimuli to explore the potential association between depression and fear. This study aimed at investigating brain alterations in a rodent model of depression when innate fear was induced in the form of the predator odor trimethylthiazoline (TMT). Flinders sensitive line (FSL) rats, a genetic animal model of depression, and their control counterpart Flinders resistant
Multiple Genetic Elements Carry the Tetracycline Resistance Gene tet(W) in the Animal Pathogen Arcanobacterium pyogenes
Stephen J. Billington; B. Helen Jost
The tet(W) gene is associated with tetracycline resistance in a wide range of bacterial species, including obligately anaerobic rumen bacteria and isolates from the human gut and oral mucosa. However, little is known about how this gene is disseminated and the types of genetic elements it is carried on. We examined tetracycline-resistant isolates of the animal commensal and opportunistic pathogen
Facile patterning of genetically engineered M13 bacteriophage for directional growth of human fibroblast cells
Lee, Seung-Wuk
Facile patterning of genetically engineered M13 bacteriophage for directional growth of human developed a facile methodology to pattern engineered phage for directional guidance of human fibroblasts of cells that utilizes nanofibrous RGD-engineered phages in conjunction with micro- contact printing
Genetically modified animals from life-science, socio-economic and ethical perspectives: examining issues in an EU policy context.
Frewer, L J; Kleter, G A; Brennan, M; Coles, D; Fischer, A R H; Houdebine, L M; Mora, C; Millar, K; Salter, B
The interdisciplinary EC consortium (the PEGASUS project) aimed to examine the issues raised by the development, implementation and commercialisation of genetically modified (GM) animals, and derivative foods and pharmaceutical products. The results integrated existing social (including existing public perception) environmental and economic knowledge regarding GM animals to formulate policy recommendations relevant to new developments and applications. The use of GM in farmed animals (aquatic, terrestrial and pharmaceutical) was mapped and reviewed. A foresight exercise was conducted to identity future developments. Three case studies (aquatic, terrestrial and pharmaceutical) were applied to identify the issues raised, including the potential risks and benefits of GM animals from the perspectives of the production chain (economics and agri-food sector) and the life sciences (human and animal health, environmental impact, animal welfare and sustainable production). Ethical and policy concerns were examined through application of combined ethical matrix method and policy workshops. The case studies were also used to demonstrate the utility of public engagement in the policy process. The results suggest that public perceptions, ethical issues, the competitiveness of EU animal production and risk-benefit assessments that consider human and animal health, environmental impact and sustainable production need to be considered in EU policy development. Few issues were raised with application in the pharmaceutical sector, assuming ethical and economic issues were addressed in policy, but the introduction of agricultural GM animal applications should be considered on a case-by-case basis. PMID:23567982
NSDL National Science Digital Library
Lee, Amy.
The past few years have seen many changes in the field of genetics, including the ability to genetically clone mammals, first achieved in 1997 with a sheep named Dolly. Still a relatively new phenomenon, news stories are continually detailing new advances in cloning, reasons why cloning is important, and concerns about the safety and ethics of cloning. This week's Topic In Depth highlights some recent news articles and Web sites that address the topic of animal cloning. The first site is a recent article from the Washington Post about the sheep named Dolly, the world's first cloned mammal, who has developed arthritis at a relatively young age and has caused some to question whether cloning can have adverse health effects. An ABC news.com article details the recent birth of five cloned piglets whose parent had been genetically engineered to remove a gene that causes human bodies to reject transplanted animal organs. An Associated Press article discusses some concerns raised by scientists and ethicists surrounding the idea of xenotransplantation (animal to human transplantation). For users who need a primer on what exactly cloning means and why it is done, check out the Cloning Fact Sheet. Developed by the Human Genome Project, it provides short, non-technical explanations of the different types of cloning and some links to other cloning related Web sites. Those users looking for more detailed information about cloning technology will find the next two sites interesting. PPL Therapeutics, which created the five piglets and collaborated with the Roslin Institute to clone Dolly, provides news articles and technical descriptions of cloning and related genetic technology. The Pharmaceutical Research and Manufacturers of America's Web site provides links to a tremendous amount of information surrounding all aspects of cloning, including recent congressional activity, news, and general resources. Although focused more heavily on human cloning, The American Journal of Bioethics Online has a Web page with links to various articles relating to the ethical issues involved with cloning and genetics.
Skin cleaning with kerosene facilitates passage of carcinogens to the lungs of animals treated with used gasoline engine oil.
Lee, J H; Roh, J H; Burks, D; Warshawsky, D; Talaska, G
Solvents such as kerosene or gasoline may be used by workers to clean their skin following contact with oily materials. This practice is not recommended, as it is well known that the solvent will defat the skin. Many also suspect that solvent washing may increase exposure by carrying materials through the skin; however, there is little documentation of this. Auto mechanics may be exposed to used gasoline engine oil (UGEO), an animal carcinogen which forms carcinogen-DNA adducts in skin and lung following topical application. This study was designed to determine if cleaning with kerosene following exposure to UGEO altered absorption of carcinogens from this material. UGEO or new oil (NO) was applied to the shaved skins of groups of HSD-ICR mice for five days. At 1 or 8 hours after application, the treated skins were cleaned with either kerosene or a commercial cleaner, or were not cleaned. Animals were sacrificed 24 hours after the last application, skins and lungs harvested, and DNA analyzed for carcinogen-DNA adducts by 32P-postlabeling. Five applications of UGEO significantly increased carcinogen-DNA adduct levels in both lungs and skin compared to animals treated with NO. DNA adduct levels in the skin were reduced significantly in groups washed with kerosene or commercial cleaner. Washing at one as opposed to eight hours after UGEO application resulted in lower adduct levels regardless of cleaner. DNA adduct levels in the lung were reduced when the commercial cleaner was used, again in a time-related fashion. However, cleaning with kerosene resulted in mean carcinogen-DNA adduct levels in the lung which were significantly higher than even the positive controls, regardless of cleaning time. This is the first demonstration that kerosene cleaning facilitates passage of carcinogens through the skin, resulting in higher levels of genetic damage in a critical internal organ. PMID:10750280
The feasibility of ureteral tissue engineering using autologous veins: an orthotopic animal model with long term results
Background In an earlier study we demonstrated the feasibility to create tissue engineered venous scaffolds in vitro and in vivo. In this study we investigated the use of tissue engineered constructs for ureteral replacement in a long term orthotopic minipig model. In many different projects well functional ureretal tissue was established using tissue engineering in animals with short-time follow up (12Āweeks). Therefore urothelial cells were harvested from the bladder, cultured, expanded in vitro, labelled with fluorescence and seeded onto the autologous veins, which were harvested from animals during a second surgery. Three days after cell seeding the right ureter was replaced with the cell-seeded matrices in six animals, while further 6 animals received an unseeded vein for ureteral replacement. The animals were sacrificed 12, 24, and 48Āweeks after implantation. Gross examination, intravenous pyelogram (IVP), H&E staining, Trichrome MassonÆs Staining, and immunohistochemistry with pancytokeratin AE1/AE3, smooth muscle alpha actin, and von Willebrand factor were performed in retrieved specimens. Results The IVP and gross examination demonstrated that no animals with tissue engineered ureters and all animals of the control group presented with hydronephrosis after 12Āweeks. In the 24-week group, one tissue engineered and one unseeded vein revealed hydronephrosis. After 48Āweeks all tissue engineered animals and none of the control group showed hydronephrosis on the treated side. Histochemistry and immunohistochemistry revealed a multilayer of urothelial cells attached to the seeded venous grafts. Conclusions Venous grafts may be a potential source for ureteral reconstruction. The results of so far published ureteral tissue engineering projects reveal data up to 12Āweeks after implantation. Even if the animal numbers of this study are small, there is an increasing rate of hydronephrosis revealing failure of ureteral tissue engineering with autologous matrices in time points longer than 3Āmonths after implantation. Further investigations have to prove adequate clinical outcome and appropriate functional long-term results. PMID:25381044
Monitoring photodynamic therapy of localized infections by bioluminescence imaging of genetically engineered bacteria
Demidova, Tatiana N; Gad, Faten; Zahra, Touqir; Francis, Kevin P; Hamblin, Michael R
The increasing occurrence of multi-antibiotic resistant microbes has led to the search for alternative methods of killing pathogens and treating infections. Photodynamic therapy (PDT) uses the combination of non-toxic dyes and harmless visible light to produce reactive oxygen species that can kill mammalian and microbial cells. Although the photodynamic inactivation of bacteria has been known for over a hundred years, its use to treat infections has not been much developed. This may be partly due to the difficulty of monitoring the effectiveness of PDT in animal models of infection. In order to facilitate this monitoring process, we have developed a procedure that uses bioluminescent genetically engineered bacteria and a light sensitive imaging system to allow real-time visualization of infections. When these bacteria are treated with PDT in vitro, the loss of luminescence parallels the loss of colony-forming ability. We have developed several models of infections in wounds and soft-tissue abscesses in mice that can be followed by bioluminescence imaging. The size and intensity of the infection can be sequentially monitored in a non-invasive fashion in individual mice in real-time. When photosensitizers are introduced into the infected tissue followed by illumination with red light, a light-dose dependent loss of luminescence is seen. If the bacterium is invasive, the loss of luminescence correlates with increased survival of the mice, whilst animals in control groups die of sepsis within five days. Healing of the PDT treated wounds is not impaired and may actually be improved. This approach can allow many animal models of localized infections to be accurately monitored for efficacy of treatment by PDT. PMID:16040251
Genetically Engineered Transvestites Reveal Novel Mating Genes in Budding Yeast
Huberman, Lori B.; Murray, Andrew W.
Haploid budding yeast has two mating types, defined by the alleles of the MAT locus, MATa and MAT?. Two haploid cells of opposite mating types mate by signaling to each other using reciprocal pheromones and receptors, polarizing and growing toward each other, and eventually fusing to form a single diploid cell. The pheromones and receptors are necessary and sufficient to define a mating type, but other mating-type-specific proteins make mating more efficient. We examined the role of these proteins by genetically engineering ōtransvestiteö cells that swap the pheromone, pheromone receptor, and pheromone processing factors of one mating type for another. These cells mate with each other, but their mating is inefficient. By characterizing their mating defects and examining their transcriptomes, we found Afb1 (a-factor barrier), a novel MAT?-specific protein that interferes with a-factor, the pheromone secreted by MATa cells. Strong pheromone secretion is essential for efficient mating, and the weak mating of transvestites can be improved by boosting their pheromone production. Synthetic biology can characterize the factors that control efficiency in biological processes. In yeast, selection for increased mating efficiency is likely to have continually boosted pheromone levels and the ability to discriminate between partners who make more and less pheromone. This discrimination comes at a cost: weak mating in situations where all potential partners make less pheromone. PMID:24121774
Selective detection of gold using genetically engineered bacterial reporters.
Cerminati, Sebastißn; Soncini, Fernando C; Checa, Susana K
Salmonella typhimurium harbours a Au-resistance system whose expression is controlled by GolS, a transcriptional regulator of the MerR family that selectively detects Au with high sensitivity. We developed both Salmonella and genetically engineered Escherichia coli strains as Au-selective whole-cell biosensors by coupling the strictly regulated GolS-dependent golB promoter to the gfp reporter gene. The bio-reporters were evaluated under different laboratory conditions and calibrated for their use as selective Au detectors. Due to the intrinsic characteristics of the regulatory protein, the transgenic E. coli sensor exhibits low background, high signal-to-noise ratio, and improved sensitivity for detection of Au ions in a wide range of concentrations (up to 470 nM) with a calculated detection limit of ?33 nM (6 Ąg?L(-1) or parts per billion) Au(I). The fluorescent Au-sensing bacteria exhibit also minimal interference by chemically related metals such as Cu or Ag that are commonly found in Au deposits. These highly specific and sensitive Au detectors might allow the development of rapid and robust screening tools to improve discovery and extraction procedures. PMID:21618467
Genetic engineering of yellow betalain pigments beyond the species barrier
Nakatsuka, Takashi; Yamada, Eri; Takahashi, Hideyuki; Imamura, Tomohiro; Suzuki, Mariko; Ozeki, Yoshihiro; Tsujimura, Ikuko; Saito, Misa; Sakamoto, Yuichi; Sasaki, Nobuhiro; Nishihara, Masahiro
Betalains are one of the major plant pigment groups found in some higher plants and higher fungi. They are not produced naturally in any plant species outside of the order Caryophyllales, nor are they produced by anthocyanin-accumulating Caryophyllales. Here, we attempted to reconstruct the betalain biosynthetic pathway as a self-contained system in an anthocyanin-producing plant species. The combined expressions of a tyrosinase gene from shiitake mushroom and a DOPA 4,5-dioxygenase gene from the four-o'clock plant resulted in successful betalain production in cultured cells of tobacco BY2 and Arabidopsis T87. Transgenic tobacco BY2 cells were bright yellow because of the accumulation of betaxanthins. LC-TOF-MS analyses showed that proline-betaxanthin (Pro-Bx) accumulated as the major betaxanthin in these transgenic BY2 cells. Transgenic Arabidopsis T87 cells also produced betaxanthins, but produced lower levels than transgenic BY2 cells. These results illustrate the success of a novel genetic engineering strategy for betalain biosynthesis. PMID:23760173
Distributed classifier based on genetically engineered bacterial cell cultures.
Didovyk, Andriy; Kanakov, Oleg I; Ivanchenko, Mikhail V; Hasty, Jeff; Huerta, Ram¾n; Tsimring, Lev
We describe a conceptual design of a distributed classifier formed by a population of genetically engineered microbial cells. The central idea is to create a complex classifier from a population of weak or simple classifiers. We create a master population of cells with randomized synthetic biosensor circuits that have a broad range of sensitivities toward chemical signals of interest that form the input vectors subject to classification. The randomized sensitivities are achieved by constructing a library of synthetic gene circuits with randomized control sequences (e.g., ribosome-binding sites) in the front element. The training procedure consists in reshaping of the master population in such a way that it collectively responds to the "positive" patterns of input signals by producing above-threshold output (e.g., fluorescent signal), and below-threshold output in case of the "negative" patterns. The population reshaping is achieved by presenting sequential examples and pruning the population using either graded selection/counterselection or by fluorescence-activated cell sorting (FACS). We demonstrate the feasibility of experimental implementation of such system computationally using a realistic model of the synthetic sensing gene circuits. PMID:25349924
Gene flow in genetically engineered perennial grasses: Lessons for modification of dedicated bioenergy crops
The potential ecological consequences of the commercialization of genetically engineered (GD) crops have been the subject of intense debate, particularly when the GE crops are perennial and capable of outcrossing to wild relatives. The essential ecological impact issues for engi...
The establishment of genetically engineered canola populations in the U.S.
Concerns regarding the commercial release of genetically engineered (GE) crops include naturalization, introgression to sexually compatible relatives and the transfer of beneficial traits to native and weedy species through hybridization. To date there have been few documented re...
Plasmid pSI30 was constructed to increase the sensitivity of detection of a genetically engineered microorganism (GEM) and its recombinant DNA in environmental samples. his broad host-range, mobilizable plasmid contained chlorocatechol (clc) degradative genes, antibiotic resistan...
Human Nature Genetically Re-Engineered: Moral Responsibilities to Future Generations
H. Tristram Engelhardt
\\u000a The prospect of human genetic germ-line engineering raises questions regarding the propriety of altering the human genome.\\u000a It raises questions as well regarding the ways in which one might understand responsibilities to the future generations who\\u000a will experience the result of such alterations. This essay explores the difficulty of disclosing content-full obligations\\u000a regarding genetic germ-line engineering. Instead, as this essay
The regulation of genetically engineered marine organisms released into the coastal environment: an exploratory analysisĀ
MacGregor, Carol Lea
model for assessing the risks of releasing genetically engineered organisms into the marine environment. The current federal regulatory scheme is analyzed against the proposed model involving the following steps: 1) hazard identification; 2) risk... characterization; 3) exposure assessment; 4) hazard evaluation; and 5) risk analysis. The legal implications of a standard methodology for assessing the risks of releasing genetically engineered organisms are presented. Over the past ten years, biotechnology...
Small-scale field test of the genetically engineered lacZY marker
H. A. Hattemer-Frey; E. J. Brandt; C. C. Travis
Commercial genetic engineering is advancing into areas that require the small-scale introduction of genetically engineered microorganisms (GEMs) to better quantify variables that affect microorganism distribution and survival and to document potential long-term consequences. A recombinant DNA marker system, the lacZY marker, developed by the Monsanto Agricultural Co., enables the distribution and fate of marked fluorescent pseudomonad organisms to be monitored
Tumor-specific gene delivery using genetically engineered bacteria.
Theys, J; Barbķ, S; Landuyt, W; Nuyts, S; Van Mellaert, L; Wouters, B; Annķ, J; Lambin, P
The loco-regional control of cancer remains a major contributor to the treatment outcome for many cancer patients prescribed conventional radiotherapy or chemotherapy. Failure of treatment coupled with the realisation that cancer is essentially a genetic disease has led to development of many clinical protocols based on gene therapy. In this review, we will describe an alternative gene delivery system based on the use of non-pathogenic bacteria. Tumor regressions have been reported long ago in patients with bacterially infected tumors, suggesting that bacteria could target tumors and have local anti-tumor effects. The basis of this phenomenon is attributable to the unique properties of the tumor micro-environment. The presence of hypoxic and/or necrotic areas provides a haven for a number of anaerobic bacteria and over the past 60 years, several strains of anaerobic bacteria have been shown to localise within and cause cell lysis of experimental animal tumors. One of the most important strains in that context is Clostridium. Other bacteria have also been implicated in experimental anti-cancer settings. Of these, attenuated Salmonella strains capable of both selective amplification within tumors and expression of effector genes encoding therapeutic proteins are probably the most promising. We will discuss the potential advantages and the pitfalls of this alternative delivery approach. We will emphasize the importance of hypoxia in solid tumors and discuss the potential of radiation-inducible promoters and combined treatment modalities, involving vascular targeting and radiotherapy. We believe that this approach will act in a complementary way to current radiotherapy and chemotherapy treatments of solid tumors. PMID:12762480
468 IEEE TRANSACTIONS ON ENGINEERING MANAGEMENT, VOL. 56, NO. 3, AUGUST 2009 The Effects of Moving Animation on Recall, Hedonic
Hui, Kai-Lung
468 IEEE TRANSACTIONS ON ENGINEERING MANAGEMENT, VOL. 56, NO. 3, AUGUST 2009 The Effects of Moving Animation on Recall, Hedonic and Utilitarian Perceptions, and Attitude Yee-Lin Lai, Kevin K. Y. Kuan, Kai-Lung Hui, and Na Liu Abstract--An increasing number of firms are investing in mov- ing animation to create
Short communication: Validation of two animal models for estimation of genetic trends for female fertility in Norwegian dairy cattle.
Andersen-Ranberg, I M; Klemetsdal, G; Heringstad, B
Two animal models were compared with respect to potential bias in genetic trend estimates for female fertility and for their predictive ability. In addition to either a fixed effect for month of first insemination or for month-year of first insemination, the models had fixed effects of age and double insemination and random effects of herd-year and animal. The model with a fixed effect of month of first insemination had a larger positive genetic trend for 56-d nonreturn rate in virgin heifers (0.16% yr), smaller downward bias, and somewhat higher predictive ability. These results demonstrate the importance of verifying models to be used in the calculation of breeding values. PMID:14740849
Payments for agrobiodiversity conservation services for sustained on-farm utilization of plant and animal genetic resources
Ulf Narloch; Adam G. Drucker; Unai Pascual
This paper discusses the potential application of Payment for Ecosystem Services-like schemes to tackle market failures associated with the public good characteristics of agrobiodiversity conservation services. So called payments for agrobiodiversity conservation services (PACS) would increase the private benefits from utilizing local plant and animal genetic resources on-farm through voluntary reward mechanisms, so as to sustain their on-farm conservation. Theoretical
Recombineering: in vivo genetic engineering in E. coli, S. enterica, and beyond.
Sawitzke, James A; Thomason, Lynn C; Costantino, Nina; Bubunenko, Mikhail; Datta, Simanti; Court, Donald L
"Recombineering," in vivo genetic engineering with short DNA homologies, is changing how constructs are made. The methods are simple, precise, efficient, rapid, and inexpensive. Complicated genetic constructs that can be difficult or even impossible to make with in vitro genetic engineering can be created in days with recombineering. DNA molecules that are too large to manipulate with classical techniques are amenable to recombineering. This technology utilizes the phage lambda homologous recombination functions, proteins that can efficiently catalyze recombination between short homologies. Recombineering can be accomplished with linear PCR products or even single-stranded oligos. In this chapter we discuss methods of and ways to use recombineering. PMID:17352923
Withdrawal emotional-regulation in infant rats from genetic animal models of depression.
Braw, Y; Malkesman, O; Merenlender, A; Bercovich, A; Dagan, M; Overstreet, D H; Weller, A
Children of depressed parents exhibit high rates of emotion-dysregulation, characterized by excessive withdrawal or approach strategies toward the mother in infancy. The understanding of factors affecting the establishment of these behavioral deficits is limited. The current study utilized two genetic animal models of depression, the Wistar-Kyoto (WKY) and Flinders Sensitive Line (FSL) rat strains. In addition, in order to assess the interactive effects of depressive vulnerability and exposure to early life stress, the subjects were raised either in a standard rearing condition or exposed to mild chronic-stress on postnatal days (PND) 2-9. On PND 10-11, an isolation test examined the pups' emotion-regulation. WKY pups produced less separation-induced ultrasonic vocalizations (USV) and proximity-seeking behaviors, compared to controls. In addition, WKY pups did not show the expected potentiation effect that was evident in control pups (an increase in USV and pivoting behavior after a short reunion with the dam). FSL pups exhibited less proximity-seeking behaviors compared to controls while showing levels of USV, potentiation of USV, and change in proximity-seeking behaviors that were similar to controls. No differences between the strains were found in self-grooming. The early life chronic-stress paradigm had no effect on the behaviors of the pups, indicating either stress-resilience or a limited effect of the paradigm. Overall, the results tentatively imply a tendency of the WKY and FSL pups towards withdrawal behavior instead of approach-behavior when regulating emotion, with a more pronounced pattern in WKY pups. This behavioral profile is reminiscent of avoidant attachment, a characteristic of many children of depressed parents. PMID:18539346
Establishment of the mathematical model for diagnosing the engine valve faults by genetic
Fernandez, Thomas
1 Establishment of the mathematical model for diagnosing the engine valve faults by genetic of the proposed strategy is validated by an illustrative example, in which three kinds of valve states inherent in a six- cylinders/four-stroke cycle diesel engine, i.e. normal condition, valve-tappet clearance and gas
Genetically engineered bacteria: An emerging tool for environmental remediation and future research perspectives
Jay Shankar Singh; P. C. Abhilash; H. B. Singh; Rana P. Singh; D. P. Singh
This minireview explores the environmental bioremediation mediated by genetically engineered (GE) bacteria and it also highlights the limitations and challenges associated with the release of engineered bacteria in field conditions. Application of GE bacteria based remediation of various heavy metal pollutants is in the forefront due to eco-friendly and lesser health hazards compared to physico-chemical based strategies, which are less
The basis for the first U.S. Environmental Protection Agency approval for the field release of a genetically-engineered virus has been to take advantage of the biological properties of baculovirus in such a way that an engineered virus could possess enhanced pesticidal properties...
From microcarriers to hydrodynamics: introducing engineering science into animal cell culture.
Croughan, Matthew S; Hu, Wei-Shou
Professor Daniel I.C. Wang has conducted research in animal cell culture for approximately 40 years. Over that long time period and still to this day, he successfully addresses a multitude of engineering challenges, taking a unique, creative, systems-driven but still fundamental approach. As mammalian cell culture has become the predominant method of manufacturing therapeutic proteins, the impact of his leadership, not only in research but also student recruitment and education, has played a key role in the success of the bio/pharmaceutical industry. PMID:16933297
Use of transgenic animals in toxicology
David J Kirkland
Transgenic and genetically engineered animals are being increasingly used in the study of diseases and for safety assessments for new products. There are four main areas in which they influence pharmaceutical development; two of these, new disease models and `humanized' animals for the assessment of biopharmaceuticals, have not yet made their impact upon toxicology and so will only be briefly
Genetically engineered structure-based vaccine for bluetongue disease.
Roy, P
At present the only vaccines used against bluetongue (BT) disease are live-attenuated virus vaccines. Since viruses with RNA genomes such as BT virus (BTV) have a high frequency of mutations, live virus vaccines could have breakthroughs (vaccine failures) and mutate into virulent strains. In addition, multiple BTV serotypes exist in nature which could potentially cause additional problems with live virus vaccines. The BTV genome is made up of 10 segments and therefore potentially could exchange these segments (or genes) randomly between different viruses including vaccine strains, generating novel viruses with mixed genes. Hence it is necessary to develop BTV vaccines that pose no such threat. Ideally BTV vaccines should be completely devoid of harmful genes. Recent protein expression technology has provided novel approaches for the development of intrinsically safe vaccines. The technology involves the synthesis of immunogenic proteins and particles that elicit highly protective immune responses. We have generated such vaccines, termed virus-like particle (VLP) vaccines. These vaccines (which do not carry either BTV or foreign genes) give the immune system information about viral structures so that it can generate a complete defence against the real virus infection very efficiently. A series of vaccine trials were undertaken outdoors under natural UV light using 50-200 BTV-susceptible sheep per trial. Vaccination trials of sheep showed that the VLPs were highly immunogenic, and protected sheep when animals were challenged with virulent virus even 15 months after the first immunisation. Moreover, a cocktail of five VLPs afforded protection against not only each of the homologous BTV serotypes but also against certain heterologous serotypes that are genetically related to some of these vaccine strains. VLPs representing a number of serotypes are currently available and can be produced fairly quickly if there is such a need. Based on our sequence data it can be predicted that a mixture of seven or eight types of VLPs (that are already available) will provide protection against at least 10 or more serotypes depending on their phylogenetic relationships. BTV VLPs offer particular advantages as potential vaccines over other systems. Large quantities of VLPs can be produced and easily purified using a one-step protocol based on the physical properties of the particle. More importantly, these particles are devoid of any nucleic acid and thus pose no threat by re-assortment or the re-emergence of virulence that attenuated vaccines can cause. PMID:20422594
The facial animation engine: toward a high-level interface for the design of MPEG4 compliant animated faces
Fabio Lavagetto; Roberto Pockaj
We propose a method for implementing a high-level interface for the synthesis and animation of animated virtual faces that is in full compliance with MPEG-4 specifications. This method allows us to implement the simple facial object profile and part of the calibration facial object profile. In fact, starting from a facial wireframe and from a set of configuration files, the
NSDL National Science Digital Library
National Science Teachers Association (NSTA)
What affects how physical characteristics are transmitted from parent to offspring? This is a question that can be answered at many levels. Molecular biologists examine the pattern of nucleotides in deoxyribonucleic acid (DNA) and the effect of mutations on the proteins produced. Classical geneticists explore the patterns by which traits are transmitted through families. Medical geneticists attempt to describe and develop treatments for diseases that have a genetic component. Genetic engineers analyze how traits can be altered in organisms through modern technology. These are only a few of the strategies that scientists employ to explain the nature of heredity. Explore historical perspectives on the study of genetics and investigate how cutting-edge technology is being used to expand our understanding of heredity.
On-chip whole-animal manipulation for high-throughput subcellular-resolution in-vivo drug/genetic screening
Yanik, Mehmet Fatih
Techniques for rapid and automated small-animal manipulation and immobilization are necessary for high-throughput in vivo genetic/drug screens using cellular and sub-cellular features in multicellular organisms. We present ...
Vicariance and dispersal across an intermittent barrier: population genetic structure of marine animals across the Torres Strait land bridge
NASA Astrophysics Data System (ADS)
Mirams, A. G. K.; Treml, E. A.; Shields, J. L.; Liggins, L.; Riginos, C.
Biogeographic barriers, some transitory in duration, are likely to have been important contributing factors to modern marine biodiversity in the Indo-Pacific region. One such barrier was the Torres Strait land bridge between continental Australia and New Guinea that persisted through much of the late Pleistocene and separated Indian and Pacific Ocean taxa. Here, we examine the patterns of mitochondrial DNA diversity for marine animals with present-day distributions spanning the Torres Strait. Specifically, we investigate whether there are concordant signatures across species, consistent with either vicariance or recent colonization from either ocean basin. We survey four species of reef fishes ( Apogon doederleini, Pomacentrus coelestis, Dascyllus trimaculatus, and Acanthurus triostegus) for mtDNA cytochrome oxidase 1 and control region variation and contrast these results to previous mtDNA studies in diverse marine animals with similar distributions. We find substantial genetic partitioning (estimated from F-statistics and coalescent approaches) between Indian and Pacific Ocean populations for many species, consistent with regional persistence through the late Pleistocene in both ocean basins. The species-specific estimates of genetic divergence, however, vary greatly and for reef fishes we estimate substantially different divergence times among species. It is likely that Indian and Pacific Ocean populations have been isolated for multiple glacial cycles for some species, whereas for other species genetic connections have been more recent. Regional estimates of genetic diversity and directionality of gene flow also vary among species. Thus, there is no apparent consistency among historical patterns across the Torres Strait for these co-distributed marine animals.
Coppola, Antonietta; Moshķ, Solomon L
Epilepsy accounts for a significant portion of the dis-ease burden worldwide. Research in this field is fundamental and mandatory. Animal models have played, and still play, a substantial role in understanding the patho-physiology and treatment of human epilepsies. A large number and variety of approaches are available, and they have been applied to many animals. In this chapter the in vitro and in vivo animal models are discussed,with major emphasis on the in vivo studies. Models have used phylogenetically different animals - from worms to monkeys. Our attention has been dedicated mainly to rodents.In clinical practice, developmental aspects of epilepsy often differ from those in adults. Animal models have often helped to clarify these differences. In this chapter, developmental aspects have been emphasized.Electrical stimulation and chemical-induced models of seizures have been described first, as they represent the oldest and most common models. Among these models, kindling raised great interest, especially for the study of the epileptogenesis. Acquired focal models mimic seizures and occasionally epilepsies secondary to abnormal cortical development, hypoxia, trauma, and hemorrhage.Better knowledge of epileptic syndromes will help to create new animal models. To date, absence epilepsy is one of the most common and (often) benign forms of epilepsy. There are several models, including acute pharmacological models (PTZ, penicillin, THIP, GBL) and chronic models (GAERS, WAG/Rij). Although atypical absence seizures are less benign, thus needing more investigation, only two models are so far available (AY-9944,MAM-AY). Infantile spasms are an early childhood encephalopathy that is usually associated with a poor out-come. The investigation of this syndrome in animal models is recent and fascinating. Different approaches have been used including genetic (Down syndrome,ARX mutation) and acquired (multiple hit, TTX, CRH,betamethasone-NMDA) models.An entire section has been dedicated to genetic models, from the older models obtained with spontaneous mutations (GEPRs) to the new engineered knockout, knocking, and transgenic models. Some of these models have been created based on recently recognized patho-genesis such as benign familial neonatal epilepsy, early infantile encephalopathy with suppression bursts, severe myoclonic epilepsy of infancy, the tuberous sclerosis model, and the progressive myoclonic epilepsy. The contribution of animal models to epilepsy re-search is unquestionable. The development of further strategies is necessary to find novel strategies to cure epileptic patients, and optimistically to allow scientists first and clinicians subsequently to prevent epilepsy and its consequences. PMID:22938964
Integration of optimization by genetic algorithms into an L-system-based animation system
Hansrudi Noser; Peter Stucki; Hans-Peter Walser
In computer graphics L-systems represent a powerful rule-based language for modeling complex objects and their animation. However, designing objects and animations by rules is a difficult task because designers often cannot foresee the consequences of rules. This is especially true for non-experts in the domain. Therefore, we propose to enhance an L-system based animation system with evolutionary features based on
Genetic engineering: a matter that requires further refinement in Spanish secondary school textbooks
NASA Astrophysics Data System (ADS)
MartĒnez-Gracia, M. V.; Gil-Qu²lez, M. J.
Genetic engineering is now an integral part of many high school textbooks but little work has been done to assess whether it is being properly addressed. A checklist with 19 items was used to analyze how genetic engineering is presented in biology textbooks commonly used in Spanish high schools, including the content, its relationship with fundamental genetic principles, and how it aims to improve the genetic literacy of students. The results show that genetic engineering was normally introduced without a clear reference to the universal genetic code, protein expression or the genetic material shared by all species. In most cases it was poorly defined, without a clear explanation of all the relevant processes involved. Some procedures (such as vectors) were explained in detail without considering previous student knowledge or skills. Some books emphasized applications such as the human genome project without describing DNA sequencing. All books included possible repercussions, but in most cases only fashionable topics such as human cloning. There was an excess of information that was not always well founded and hence was unsuitable to provide a meaningful understanding of DNA technology required for citizens in the twenty-first century.
Monitoring for genetically engineered pseudomonas species in monterey county
Supkoff, D.; Opgenorth, D.; Lai, C.; Segawa, R.; Koehler, D.
A field monitoring study was conducted to determine if genetically altered Pseudomonas fluorescens or P. syringae had been applied to sites in Monterey County. A series of diagnostic tests for antibiotic resistance, fluorescence ability, oxidase and arginine dihydrolase activities, hypersensitivity reaction and ice nucleation ability were conducted to screen bacteria isolated from field and control samples. No bacteria were detected from field samples which matched the expected test profiles of genetically altered bacterial products. In contrast, bacteria were consistently isolated from positive control samples with the expected characteristics of genetically altered bacteria.
Javed Akhter; Mohammed Qutub; Norman Burnham
For thousands of years, humans have taken advantage of naturally occurring genetic variation within species to selectively breed organisms with desirable traits. Many of the characteristics of domestic animals and agricultural crops have been developed through selective breeding. What is revolutionary about genetic engineering is that it involves the transfer of genetic material between organisms that would never be able
Isolates of Cryptococcus neoformans from Infected Animals Reveal Genetic Exchange in Unisexual, Ā Mating Type Populations
Tien Bui; Xiaorong Lin; Richard Malik; Joseph Heitman; Dee Carter
Sexual reproduction and genetic exchange are important for the evolution of fungal pathogens and for producing potentially infective spores. Studies to determine whether sex occurs in the pathogenic yeast Cryptococcus neoformans var. grubii have produced enigmatic results, however: basidiospores are the most likely infective propagules, and clinical isolates are fertile and genetically diverse, consistent with a sexual species, but almost
Spatial genetic structure of Simarouba amara Aubl. (Simaroubaceae), a dioecious, animal-dispersed
Bermingham, Eldredge
. amara, in comparison to other recent studies, may be explained by pollen and seed dispersal over the 50); or (3) use of genetic markers to reconstruct seed and pollen movement through parentage analyses: (1) to describe the genetic diversity of a mapped population of S. amara within a lowland Neotropical
Role of reproductive technologies and genetic resource banks in animal conservation
William V. Holt; Amanda R. Pickard
In combination with modern reproductive technologies, there is potential to use frozen and stored germplasm (genetic resource banks) to support conservation measures for the main- tenance of genetic diversity in threatened species. However, turning this idea into reality is a complex process, requiring interdisciplinary collaboration and clearly defined goals. As the number of species deserving the attention of conservation scientists
The Facial Animation Engine: towards a high-level interface for the design of MPEG4 compliant animated faces
Fabio Lavagetto; Roberto Pockaj
In this paper we propose a method for implementing a high-level interface for the synthesis and animation of animated virtual faces that is in full compliance with MPEG-4 specifications. This method allows us to implement the Simple Facial Object profile and part of the Calibration Facial Object Profile. In fact, starting from a facial wire-frame and from a set of
Moral and Legal Decisions in Reproductive and Genetic Engineering
ERIC Educational Resources Information Center
Heim, Werner G.
Discusses the moral and ethical issues raised by the imminent possibilities for genetic and reproductive manipulation of humans, the responsibilities of scientists, moralists, and social scientists, and the role of teachers in public information. (AL)
Genetically engineered murine models ¢ Contribution to our understanding of the genetics, molecular pathology and therapeutic targeting of neuroblastoma
Chesler, Louis; Weiss, William A.
Genetically engineered mouse models (GEMM) have made major contributions to a molecular understanding of several adult cancers and these results are increasingly being translated into the pre-clinical setting where GEMM will very likely make a major impact on the development of targeted therapeutics in the near future. The relationship of pediatric cancers to altered developmental programs, and their genetic simplicity relative to adult cancers provides unique opportunities for the application of new advances in GEMM technology. In neuroblastoma the well-characterized TH-MYCN GEMM is increasingly used for a variety of molecular-genetic, developmental and pre-clinical therapeutics applications. We discuss: the present and historical application of GEMM to neuroblastoma research, future opportunities, and relevant targets suitable for new GEMM strategies in neuroblastoma. We review the potential of these models to contribute both to an understanding of the developmental nature of neuroblastoma and to improved therapy for this disease. PMID:21958944
Complete Genome Sequence of Klebsiella pneumoniae Strain ATCC 43816 KPPR1, a Rifampin-Resistant Mutant Commonly Used in Animal, Genetic, and Molecular Biology Studies.
Broberg, Christopher A; Wu, Weisheng; Cavalcoli, James D; Miller, Virginia L; Bachman, Michael A
Klebsiella pneumoniae is an urgent public health threat due to the spread of carbapenem-resistant strains causing serious, and frequently fatal, infections. To facilitate genetic, molecular, and immunological studies of this pathogen, we report the complete chromosomal sequence of a genetically tractable, prototypical strain used in animal models. PMID:25291761
Genome-scale genetic engineering in Escherichia coli.
Jeong, Jaehwan; Cho, Namjin; Jung, Daehee; Bang, Duhee
Genome engineering has been developed to create useful strains for biological studies and industrial uses. However, a continuous challenge remained in the field: technical limitations in high-throughput screening and precise manipulation of strains. Today, technical improvements have made genome engineering more rapid and efficient. This review introduces recent advances in genome engineering technologies applied to Escherichia coli as well as multiplex automated genome engineering (MAGE), a recent technique proposed as a powerful toolkit due to its straightforward process, rapid experimental procedures, and highly efficient properties. PMID:23624241
Small Animal Bone Biomechanics
Vashishth, Deepak
Animal models, in particular mice, offer the possibility of naturally achieving or genetically engineering a skeletal phenotype associated with disease and conducting destructive fracture tests on bone to determine the resulting change in boneÆs mechanical properties. Several recent developments, including nano- and micro- indentation testing, microtensile and microcompressive testing, and bending tests on notched whole bone specimens, offer the possibility to mechanically probe small animal bone and investigate the effects of aging, therapeutic treatments, disease, and genetic variation. In contrast to traditional strength tests on small animal bones, fracture mechanics tests display smaller variation and therefore offer the possibility of reducing sample sizes. This article provides an analysis of what such tests measure and proposes methods to reduce errors associated with testing smaller than ideal specimens. PMID:18672104
Potential effects of animal management and genetic improvement on enteric methane emissions, emissions intensity and productivity of sheep enterprises at Cowra, Australia
D. J. Alcock; R. S. Hegarty
The efficacy of technologies to reduce enteric CH4 emissions from ruminants are typically evaluated on individual animals with little consideration of enterprise scale impacts. While impacts of the many rumen manipulations being studied are hard to anticipate, there is adequate information to assess impacts of farm management changes and potential animal genetic changes on whole farm productivity and enteric CH4
Genetic engineering and the Moral Status of Non-Human Species
Anders Melin
Genetic modification leads to several important moral issues. Up until now they have mainly been discussed from the viewpoint that only individual living beings, above all animals, are morally considerable. The standpoint that also collective entities such as species belong to the moral sphere have seldom been taken into account in a more thorough way, although it is advocated by
Standardization of functional reporter and antibiotic resistance cassettes to facilitate the genetic engineering of filamentous fungi.
Sureka, Swati; Chakravorty, Arun; Holmes, Eric C; Spassibojko, Olga; Bhatt, Nupur; Wu, Dongliang; Turgeon, B Gillian
The unique physiological properties of fungi are useful for a myriad of applications, which could greatly benefit from increased control of native pathways and introduction of recombinant genes. However, fungal genetic engineering is still limited in scope and accessibility, largely due to lack of standardization. To help standardize the genetic engineering of filamentous fungi, we created BioBricks of commonly used antibiotic resistance genes, neomycin phosphotransferase (nptII) and hygromycin phosphotransferase (hph), which confer resistance to G418 (Geneticin) and hygromycin B, respectively. Additionally, we created a BioBrick of the constitutive trpC promoter, from the tryptophan biosynthesis pathway of Aspergillus nidulans, and used it to create a composite part including the GFP gene. The functionality of these parts was demonstrated in the model fungal organism Cochliobolus heterostrophus, and as these tools are in modular BioBrick format, they can be easily used to facilitate genetic engineering of other fungal species. PMID:25524098
Malignancy-Associated Vessel Tortuosity: A Computer-Assisted, MRA Study of Choroid Plexus Carcinoma in Genetically Engineered Mice
Bullitt, Elizabeth; Wolthusen, P. Anne; Brubaker, Lauren; Lin, Weili; Zeng, Donglin; Van Dyke, Terry
Background and Purpose The ability to assess tumor malignancy and to monitor treatment response non-invasively would be of value to both clinicians and animal investigators. This report describes the MR imaging characteristics of a genetically engineered mouse model of choroid plexus carcinoma (CPC) during tumor growth and progression to malignancy. We assess the ability of vessel tortuosity measurements, as calculated from high-resolution MRA images, to detect emerging CPC cancers. Methods MR images were analyzed of 9 healthy mice and of 20 CPC mice with precancerous choroid dysplasia or with cancer over a wide range of sizes. Two vessel tortuosity measures and a measure of vessel density (vessel count) were calculated from MRA images. Malignancy assessment was based upon a statistical analysis of vessel tortuosity, using an equation derived from an earlier study of human brain tumor patients. Results Choroid dysplasia was correctly judged non-malignant. On the basis of vessel count, neo-angiogenesis could not be detected until cancers were full-blown and had reached a volume of approximately 80mm3. Vessel tortuosity measurements, however, correctly identified emerging malignancy in lesions larger than 0.3mm3. Conclusion This report provides the first description of in vivo, MR imaging characteristics of genetically engineered CPC mice during the progression from dysplasia to cancer. Vessel tortuosity measurements offer promise of correctly defining even tiny tumors as malignant. PMID:16552004
Genetic engineering of flavonoid pigments to modify flower color in floricultural plants
Masahiro NishiharaTakashi Nakatsuka; Takashi Nakatsuka
Recent advances in genetic transformation techniques enable the production of desirable and novel flower colors in some important\\u000a floricultural plants. Genetic engineering of novel flower colors is now a practical technology as typified by commercialization\\u000a of a transgenic blue rose and blue carnation. Many researchers exploit knowledge of flavonoid biosynthesis effectively to\\u000a obtain unique flower colors. So far, the main
Andres, Andrew
double-stranded DNA - Recognition sties - Often generate - Molecules cut #12;4 b. Cloning vectors - Purpose of a vector = - Works because - Key considerations i. Must have ii. Usually engineered iii. Usually engineered - Vectors are limited #12;5 c. Specialized cloning vectors - Vectors are used i
Aerosol survival of a genetically engineered strain of Escherichia coli demonstrated a more rapid die-off (i.e., death rate) compared to its parental wildtype. p to 77% of a freeze-dried and air-exposed genetically engineered microorganism (GEM) and wildtype bacteria could be res...
Cellular computation and communications using engineered genetic regulatory networks
Weiss, Ron, 1970-
In this thesis, I present an engineering discipline for obtaining complex, predictable, and reliable cell behaviors by embedding biochemical logic circuits and programmed intercellular communications into cells. To accomplish ...
Systematic evaluation of a tissue-engineered bone for maxillary sinus augmentation in large animal canine model
Shaoyi Wang; Zhiyuan Zhang; Lunguo Xia; Jun Zhao; Xiaojuan Sun; Xiuli Zhang; Dongxia Ye; Hasan Uluda?; Xinquan Jiang
The objective of this study is to systematically evaluate the effects of a tissue-engineered bone complex for maxillary sinus augmentation in a canine model. Twelve sinus floor augmentation surgeries in 6 animals were performed bilaterally and randomly repaired with the following 3 groups of grafts: group A consisted of tissue-engineered osteoblasts\\/beta-TCP complex (n=4); group B consisted of beta-TCP alone (n=4);
IGERT- Genetic Engineering and Society: The Case of Transgenic Pests
Fred Gould
SUMMARY This IGERT project will create a transformative graduate education program that broadly trains students in technologies needed for manipulating pest genomes as well as methods needed to assess the environmental and social appropriateness of specific products of these manipulations. The concept of genetically manipulating a pest species to destroy or render it benign dates back to the 1940's, and
Genetically Engineered Grapevines Carrying GFLV Coat Protein and Antifreeze Genes
Georgy P. GUTORANOV; Ivan J. TSVETKOV; Violeta M. COLOVA-TSOLOVA; Atanas I. ATANASSOV
SUMMARY Biotic and abiotic stress has a negative effect on both the quality and quantity of grape production. Like many woody crops, grape has been relatively recalcitrant to in vitro manipulations. The crucial point in the process of genetic transformation is to have cells that are able to both regenerate and be transformed. A regeneration system seems to be a
The Human Rights of the Genetically Engineered Athlete
Andy Miah
Traditional definitions of what constitutes a human being in human rights discourse fail to include the new kinds of human beings that are emerging through genetic manipulation. The prospect of such technology and the knowledge that such alterations infringe on a number of human rights and so require further consideration, in order to be clear about their appropriateness for human
Preclinical Safety Evaluation of ASCs Engineered by FLPo/Frt-Based Hybrid Baculovirus: In Vitro and Large Animal Studies.
Li, Kuei-Chang; Chang, Yu-Han; Lin, Chin-Yu; Hwang, Shiaw-Min; Wang, Tzu-Hao; Hu, Yu-Chen
We recently developed hybrid baculovirus (BV) vectors that exploited FLPo/Frt-mediated DNA minicircle formation. Engineering of adipose-derived stem cells (ASCs) with the FLPo/Frt-based BV vectors enabled prolonged transgene expression and, after cell implantation into rabbits, ameliorated cartilage regeneration and bone repair. To translate the hybrid BV one step further toward clinical applications, here we assessed the biosafety profiles of the hybrid BV-engineered human ASCs (hASCs) in vitro and evaluated the immune responses elicited by the engineered porcine ASCs (pASCs) in large animals. We confirmed that the hybrid BV did not compromise the hASCs viability, immunosuppressive capacity, and surface characteristics. Neither did the hybrid BV cause chromosomal abnormality/transgene integration in vitro nor did it induce tumorigenicity in vivo. In the large animal study, pASCs were engineered with the hybrid BV expressing bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor (VEGF) and implanted into femoral bone defects in mini pigs. The hybrid BV-engineered pASCs enabled prolonged BMP2/VEGF expression and triggered the healing of massive segmental bone defects, while only eliciting transient antibody, cytokine, and local cellular immune responses stemming from the implantation procedure itself. These data altogether demonstrated the safety of the hybrid BV vectors for ASCs engineering and bone healing in large animals, hence implicating the potential in clinical applications. PMID:25602313
History of Oncolytic Viruses: Genesis to Genetic Engineering
Elizabeth Kelly; Stephen J Russell
Since the turn of the nineteenth century, when their existence was first recognized, viruses have attracted considerable interest as possible agents of tumor destruction. Early case reports emphasized regression of cancers during naturally acquired virus infections, providing the basis for clinical trials where body fluids containing human or animal viruses were used to transmit infections to cancer patients. Most often
A microcosm is described to evaluate and measure bacterial conjugation in the rhizosphere of barley and radish with strains of Pseudomonas cepacia. he purpose was to describe a standard method useful for evaluating the propensity of genetically engineered microorganisms (GEMs) to...
Rapid engineering of versatile molecular logic gates using heterologous genetic transcriptional modules.
Wang, Baojun; Buck, Martin
We designed and constructed versatile modular genetic logic gates in bacterial cells. These function as digital logic 1-input Buffer gate, 2-input and 3-input AND gates with one inverted input and integrate multiple chemical input signals in customised logic manners. Such rapidly engineered devices serve to achieve increased sensing signal selectivity. PMID:25062273
Development of enzymes and enzyme systems by genetic engineering to convert biomass to sugars
Technology Transfer Automated Retrieval System (TEKTRAN)
TITLE Development of Enzymes and Enzyme Systems by Genetic Engineering to Convert Biomass to Sugars ABSTRACT Plant cellulosic material is one of the most viable renewable resources for the worldÆs fuel and chemical feedstock needs. Currently ethanol derived from corn starch is the most common li...
Neural Network Model Predictive Control with Genetic Algorithm Optimization and Its Application to Turbofan Engine Starting
Yu Bo; Zhu Jihong
Turbofan engine starting is one of the most important procedures during the whole process of job, but also very complicated due to its nonlinear dynamic working procedure. Recognizing the weaknesses of predict model and traditional algorithm for rolling optimization to deal with strong nonlinear systems, this paper presents neural network model predictive control method with genetic algorithm optimization, and uses
The fate and survival of genetically engineered microbes is dependent on the survival, establishment, and growth of the microbial host, as well as on the maintenance, replication, and segregation of the recombinant plasmids within the bacterial host population. The interactions o...
Genetic engineering of embryonic stem cells via site-directed DNA recombination
Anna Norman; Mark MacInnes
This article reviews recent advances in genetic engi- neering of mammals utilizing DNA recombination tech- niques to produce targeted genome modifications. The general objective of these technologies is to discover novel gene functions via manipulation of gene expres- sion, regulation, or encoded protein sequences. The ad- vantage of gene site-directed DNA recombination is that the engineered variant remains in the
The purpose of this document is to summarize the methods and concep s that have been developed and used by the author and his colleagues to study the potential effects of genetically engineered microorganisms (GEMs) introduced, deliberately or accidently, into soil on microbemedi...
Genetic Engineering Approach to Enhance Adventitious Root Formation of Hardwood Cuttings
Zong-Ming Cheng; Wenhao Dai; Michael J. Bosela; Lori D. Osburn
Vegetative propagation by cuttings is critical to the horticultural industry, particularly the ornamental and fruit sections. This article briefly reviews the traditional approaches to improve root formation, specifically with hardwood cuttings. The focus of this review has been placed on the nontraditional strategies, including infection with Agrobacterium rhizogenes and genetic engineering with rol genes, or ro\\/B gene alone from A.
Controlled field release of a bioluminescent genetically engineered microorganism for bioremediation process monitoring and control
Steven Ripp; David E. Nivens; Yeonghee Ahn; Claudia Werner; John Jarrell; James P. Easter; Chris D. Cox; Robert S. Burlage; Gary S. Sayler
Pseudomonas fluorescens HK44 represents the first genetically engineered microorganism approved for field testing in the United States for bioremediation purposes. Strain HK44 harbors an introduced lux gene fused within a naphthalene degradative pathway, thereby allowing this recombinant microbe to bioluminescent as it degrades specific polyaromatic hydrocarbons such as naphthalene. The bioremediation process can therefore be monitored by the detection of
Pathology of genetically engineered mouse models of pancreatic exocrine cancer: Consensus report and recommendations
R. H. Hruban; N. V. Adsay; J. Albores-Saavedra; M. R. Anver; A. V. Biankin; G. P. Boivin; E. E. Furth; T. Furukawa; A. Klein; D. S. Klimstra; G. Kloppel; G. Y. Lauwers; D. S. Longnecker; J. Luttges; A. Maitra; G. J. A. Offerhaus; L. Perez-Gallego; M. Redston; D. A. Tuveson
Several diverse genetically engineered mouse models of pancreatic exocrine neoplasia have been developed. These mouse models have a spectrum of pathologic changes; however, until now, there has been no uniform nomenclature to characterize these changes. An international workshop, sponsored by The National Cancer Institute and the University of Pennsylvania, was held from December 1 to 3, 2004 with the goal
From Promises of Progress to Portents of Peril: Public Responses to Genetic Engineering
Dorothy Nelkin
In 1993, scientists at a George Washington University laboratory conducted a genetic engineering experiment that ōtwinnedö a nonviable human embryo. The purpose was to find a way to create additional embryos for In Vitro Fertilization, but major newspapers, popular magazines and talk shows covered the experiment as if it had actually yielded a cloning technology for the mass production of
912. Accelerated Chondrogenesis in Nanofiber Scaffolds Containing BMP2 Genetically Engineered Chondrocytes
Robert T. Gorsline; Jin Nam; Prasarn Tangkawattana; John Lannutti; Alicia L. Bertone
Articular cartilage injury and erosion is a common cause of joint pain often leading to osteoarthritis and permanent morbidity. This research sought to determine if genetically engineered chondrocytes can sustain chondrogenesis in a biodegradable nanofiber scaffold. We hypothesized that chondrocytes would adhere, seed, proliferate, and produce extracellular matrix proteins typical of articular cartilage within a polycaprolactone nanofiber scaffold and this
450. Novel, Injectable, Genetically Engineered Stem Cell-Based System for Anterior Spinal Fusion
Rod J. Oskouian; Gadi Pelled; Yoram Zilberman; Yamit Tal; Zulma Gazit; Dan Gazit
Spinal fusion has become a popular surgical technique used to provide segmental fixation and to maintain stability and correct deformity of the spine. Stem cell¢based therapy involving genetically engineered mesenchymal stem cells (MSCs) and bone morphogenetic protein (BMP) genes can promote bone formation and lead to spinal fusion. Exogenous control and monitoring of transgene expression in vivo is of great
Can Man Control His Biological Evolution? A Symposium on Genetic Engineering. Probabilities and Practicalities
ERIC Educational Resources Information Center
Djerassi, Carl
Manipulation of genes in human beings on a large scale is not possible under present conditions because it lacks economic potential and other attractions for industry. However, preventive'' genetic engineering may be a field for vast research in the future and will perhaps be approved by governments, parishes, people and industry. (PS)
Assessing the risks of invasion for genetically engineered plants: Acceptable evidence and reasonable doubt
Peter Kareiva
Agricultural biotechnology has generated concern over the risk of producing new invasive species or exacerbating current weed problems. In this paper, we introduce the regulatory context for assessing invasiveness of genetically engineered organisms in the United States and review the evidence presented by companies arguing for deregulation of particular transgenic crops. The context of invasion ecology is then used to
'HoneySweet' plum - a valuable genetically engineered fruit-tree cultivar and germplasm resource
Technology Transfer Automated Retrieval System (TEKTRAN)
æHoneySweetÆ is a plum variety developed through genetic engineering to be highly resistant to plum pox potyvirus (PPV), the causal agent of sharka disease, that threatens stone-fruit industries world-wide and most specifically, in Europe. Field testing for over 15 years in Europe has demonstrated ...
Milestones in chloroplast genetic engineering: an environmentally friendly era in biotechnology
Henry Daniell; Muhammad S. Khan; Lori Allison
Chloroplast genomes defied the laws of Mendelian inheritance at the dawn of plant genetics, and continue to defy the mainstream approach to biotechnology, leading the field in an environmentally friendly direction. Recent success in engineering the chloroplast genome for resistance to herbicides, insects, disease and drought, and for production of biopharmaceuticals, has opened the door to a new era in
A clinically relevant large-animal model for evaluation of tissue-engineered cardiac surgical patch materials
Jianhua Cui; Jinsheng Li; Megumi Mathison; Fernando Tondato; Stephen P. Mulkey; Connie Micko; Nicolas A. F. Chronos; Keith A. Robinson
Extracellular matrix (ECM) scaffolds may be useful as a tissue engineering approach toward myocardial regeneration in the infarcted heart. An appropriate large-animal model for testing the utility of biologically derived ECM in this application is needed. The purpose of this study was to develop such a model for optimal procedural success during and after patch implantation surgery.Myocardial infarction (MI) was
SYMPOSIUM: INFORMATION TECHNOLOGY FOR ANIMAL BREEDING PURPOSES Supply of Genetic InformationŚ Amount, Format, and Frequency
The volume and complexity of genetic information is increasing because of new traits and better models. New traits may include reproduction, health, and carcass. More comprehensive models include the test day model in dairy cattle or a growth model in beef cattle. More complex models, which may include nonadditive effects such as inbreeding and dominance, also provide additional information. The
Genetically Modified Foods: Are They a Risk to Human/Animal Health?
NSDL National Science Digital Library
Arpad Pusztai (Rowett Research Institute; )
The issue-focused, peer-reviewed article posits that genetically modified (GM) crops and food are being grown and consumed by the public, even though: there is little scientific study about their health risks, safety test technology is inadequate to assess potential harm, they can carry unpredictable toxins, and they may increase the risk of allergenic reactions.
Biosynthesis and genetic engineering of proanthocyanidins and (iso)flavonoids
Li Tian; Yongzhen Pang; Richard A. Dixon
Plant natural products have been used since ancient times as medicines and herbal remedies. Over the past two decades, the\\u000a results of population and intervention studies, or assays in animal or cell model systems, have revealed positive health beneficial\\u000a effects for various classes of phytochemicals, particularly polyphenols. The results of such studies have ignited an interest\\u000a in being able to
Anatomical Distribution and Genetic Relatedness of Antimicrobial Resistant E. coli from Healthy Companion Animals
Technology Transfer Automated Retrieval System (TEKTRAN)
Aims: Escherichia coli have been targeted for studying antimicrobial resistance in companion animals due to opportunistic infections and as a surrogate for resistance patterns in zoonotic organisms. The aim of our study examined antimicrobial resistance in E. coli isolated from various anatomical ...
The article reviews the few studies that have evaluated the survival of bacterial hosts and cloning vectors (e.g., phages) and the transfer of genetic information, by the processes of conjugation, transduction, and transformation, in aquatic and terrestrial environments and on pl...
The hermeneutic challenge of genetic engineering: Habermas and the transhumanists.
Edgar, Andrew
The purpose of this paper is to explore the impact that developments in transhumanist technologies may have upon human cultures (and thus upon the lifeworld), and to do so by exploring a potential debate between Habermas and the transhumanists. Transhumanists, such as Nick Bostrom, typically see the potential in genetic and other technologies for positively expanding and transcending human nature. In contrast, Habermas is a representative of those who are fearful of this technology, suggesting that it will compound the deleterious effects of the colonisation of the lifeworld, further constraining human autonomy and undermining the meaningfulness of the lifeworld by expanding the technological control and manipulation of humanity. It will be argued that these opposed positions are grounded in fundamentally different understandings of the consequences of scientific and technological advance. On one level, the transhumanists remain confident that the lifeworld has within it the resources necessary to find meaning and purpose in a society deeply infused by genetic technology. Habermas disagrees. On another level, the difference is articulated by Horkheimer and Adorno in Dialectic of Enlightenment, primarily by challenging what may be understood as a Baconian faith in science as a project for the domination of nature (where nature is an infinitely malleable material, to be dominated and shaped, without adverse consequences, purely for the purposes of human survival). While the transhumanists broadly embrace this faith, Habermas returns to something akin to Horkheimer and Adorno's pessimistic scepticism. PMID:19219641
Frank Mitloehner is an expert for agricultural air quality, animal-environmental interactions, and agricultural engineering. He is a Professor and Air Quality
Delany, Mary E.
Frank Mitloehner is an expert for agricultural air quality, animal-environmental interactions System". Dr. Mitloehner received his MS degree in Animal Science and Agricultural Engineering from joined the faculty in the Department of Animal Science at the University of California-Davis in 2002, Dr
Comparative genetic characterization of Listeria monocytogenes isolates from human and animal listeriosis cases
Gregory T. Jeffers; James L. Bruce; Patrick L. McDonough; Janet Scarlett; Kathryn J. Boor; Martin Wiedmann
Listeria monocytogenes isolates from human sporadic and epidemic cases (n fl 119) and from animal cases (n fl 76) were characterized by automated ribotyping and PCR-restriction fragment length polymorphism (PCR-RFLP) typing of the virulence genes actA and hly. This combination of typing methods differentiated 39 distinctive strains, each reflecting a unique combination of ribotypes, hly and actA alleles. Simpson's index
Acinetobacter sp. ADP1: an ideal model organism for genetic analysis and genome engineering.
Metzgar, David; Bacher, Jamie M; Pezo, Valķrie; Reader, John; D÷ring, Volker; Schimmel, Paul; MarliĶre, Philippe; de Crķcy-Lagard, Valķrie
Acinetobacter sp. strain ADP1 is a naturally transformable gram-negative bacterium with simple culture requirements, a prototrophic metabolism and a compact genome of 3.7 Mb which has recently been sequenced. Wild-type ADP1 can be genetically manipulated by the direct addition of linear DNA constructs to log-phase cultures. This makes it an ideal organism for the automation of complex strain construction. Here, we demonstrate the flexibility and versatility of ADP1 as a genetic model through the construction of a broad variety of mutants. These include marked and unmarked insertions and deletions, complementary replacements, chromosomal expression tags and complex combinations thereof. In the process of these constructions, we demonstrate that ADP1 can effectively express a wide variety of foreign genes including antibiotic resistance cassettes, essential metabolic genes, negatively selectable catabolic genes and even intact operons from highly divergent bacteria. All of the described mutations were achieved by the same process of splicing PCR, direct transformation of growing cultures and plating on selective media. The simplicity of these tools make genetic analysis and engineering with Acinetobacter ADP1 accessible to laboratories with minimal microbial genetics expertise and very little equipment. They are also compatible with complete automation of genetic analysis and engineering protocols. PMID:15514111
Project-oriented courses and project work can be realized even for the subject of animal genetics. Topics suited for this purpose are for instance the methods for optimizing breeding
Paris-Sud XI, Universitķ de
. RIEMENSCHNEIDER. Institute of Animal Husbandry and Breeding /Unit lor educational Research and Devetopnxent. - Department of Animal Production Agvicultuyal University Wageningen, The Netherlands. Animal Breeding withinProject-oriented courses and project work can be realized even for the subject of animal genetics
Convergence of stem cell behaviors and genetic regulation between animals and plants: insights from the Arabidopsis thaliana stomatal lineage.
Matos, Juliana L; Bergmann, Dominique C
Plants and animals are two successful, but vastly different, forms of complex multicellular life. In the 1600 million years since they shared a common unicellular ancestor, representatives of these kingdoms have had ample time to devise unique strategies for building and maintaining themselves, yet they have both developed self-renewing stem cell populations. Using the cellular behaviors and the genetic control of stomatal lineage of Arabidopsis as a focal point, we find current data suggests convergence of stem cell regulation at developmental and molecular levels. Comparative studies between evolutionary distant groups, therefore, have the power to reveal the logic behind stem cell behaviors and benefit both human regenerative medicine and plant biomass production. PMID:25184043
Genetic engineering of Periplaneta fuliginosa densovirus as an improved biopesticide
H. Jiang; J. M. Zhang; J. P. Wang; B. Yang; C. F. Liu; J. Lu; Y. Y. Hu
Summary.ĀĀThe smoky-brown cockroach (Periplaneta fuliginosa) densovirus (PfDNV) has previously shown potential in urban pest control. To improve its efficacy as a biopesticide, the\\u000a genome of PfDNV was engineered by inserting the insect-specific toxin gene BmKIT1 in the open reading frame encoding the major\\u000a structural proteins. A green fluorescent protein (GFP) marker was tagged to the BmKIT1 at its C-terminus for
Genetically engineered insertional mutagenesis in mice to model cancer: Sleeping Beauty.
Howell, Viive M; Colvin, Emily K
The ability to accurately model human cancer in mice enables in vivo examination of the biological mechanisms related to cancer initiation and progression as well as preclinical testing of new anticancer treatments and potential targets. The emergence of the genetically engineered Sleeping Beauty system of insertional mutagenesis has led to the development of a new generation of genetic mouse models of cancer and identification of novel cancer-causing genes. This chapter reviews the published cancer models of Sleeping Beauty and strategies using available strains to generate several models of cancer. PMID:25064115
Genetic threshold hypothesis of neocortical spike-and-wave discharges in the rat: An animal model of petit mal epilepsy
Vadasz, C.; Fleischer, A. [Nathan Kline Inst. for Psychiatric Research, Orangeburg, NY (United States); Carpi, D.; Jando, G. [State Univ. of New Jersey, Newark, NJ (United States)] [and others
Neocortical high-voltage spike-and-wave discharges (HVS) in the rat are an animal model of petit mal epilepsy. Genetic analysis of total duration of HVS (s/12 hr) in reciprocal F1 and F2 hybrids of F344 and BN rats indicated that the phenotypic variability of HVS cannot be explained by simple, monogenic Mendelian model. Biometrical analysis suggested the presence of additive, dominance, and sex-linked-epistatic effects, buffering maternal influence, and heterosis. High correlation was observed between average duration (s/episode) and frequency of occurrence of spike-and-wave episodes (n/12 hr) in parental and segregating generations, indicating that common genes affect both duration and frequency of the spike-and-wave pattern. We propose that both genetic and developmental - environmental factors control an underlying quantitative variable, which, above a certain threshold level, precipitates HVS discharges. These findings, together with the recent availability of rat DNA markers for total genome mapping, pave the way to the identification of genes that control the susceptibility of the brain to spike-and-wave discharges. 67 refs., 3 figs., 5 tabs.
Genetically selected Marchigian Sardinian alcohol-preferring (msP) rats: an animal model to study the neurobiology of alcoholism
Ciccocioppo, Roberto; Economidou, Daina; Cippitelli, Andrea; Cucculelli, Marino; Ubaldi, Massimo; Soverchia, Laura; Anbarasu, Lourdusami; Massi, Maurizio
The present article provides an up-to-date review that summarize almost 18 years of research in genetically selected Marchigian Sardinian alcohol-preferring (msP) rats. The results of this work demonstrate that msP rats have natural preference for ethanol characterized by a spontaneous binge-type of drinking leading to pharmacologically significant blood ethanol levels. This rat line is highly vulnerable to relapse and presentation of stimuli predictive of alcohol availability or foot-shock stress can reinstate extinguished drug-seeking up to 8 months from the last alcohol experience. The msP rat is highly sensitive to stress, shows an anxious phenotype and has depressive-like symptoms that recover following ethanol drinking. Interestingly, these animals have an up-regulated corticotrophin releasing factor (CRF) receptor 1 system. From clinical studies we learned that alcoholic patients often drink ethanol in the attempt to self-medicate from negative affective states and to search anxiety relief. We propose that msP rats represent an animal model that largely mimics that human alcoholic population that due to low ability to engage in stress-coping strategies drink ethanol as a tension relief strategy and for self-medication purposes. PMID:16961763
Genetically engineered theranostic mesenchymal stem cells for the evaluation of the anticancer efficacy of enzyme/prodrug systems.
Nouri, Faranak Salman; Wang, Xing; Hatefi, Arash
Over the past decade, various enzyme/prodrug systems such as thymidine kinase/ganciclovir (TK/GCV), yeast cytosine deaminase/5-fluorocytosine (yCD/5-FC) and nitroreductase/CB1954 (NTR/CB1954) have been used for stem cell mediated suicide gene therapy of cancer. Yet, no study has been conducted to compare and demonstrate the advantages and disadvantages of using one system over another. Knowing that each enzyme/prodrug system has its own strengths and weaknesses, we utilized mesenchymal stem cells (MSCs) as a medium to perform for the first time a comparative study that illustrated the impact of subtle differences among these systems on the therapeutic outcome. For therapeutic purposes, we first genetically modified MSCs to stably express a panel of four suicide genes including TK (TK007 and TK(SR39) mutants), yeast cytosine deaminase:uracil phosphoribosyltransferase (yCD:UPRT) and nitroreductase (NTR). Then, we evaluated the anticancer efficacies of the genetically engineered MSCs in vitro and in vivo by using SKOV3 cell line which is sensitive to all four enzyme/prodrug systems. In addition, all MSCs were engineered to stably express luciferase gene making them suitable for quantitative imaging and dose-response relationship studies in animals. Considering the limitations imposed by the prodrugs' bystander effects, our findings show that yCD:UPRT/5-FC is the most effective enzyme/prodrug system among the ones tested. Our findings also demonstrate that theranostic MSCs are a reliable medium for the side-by-side evaluation and screening of the enzyme/prodrug systems at the preclinical level. The results of this study could help scientists who utilize cell-based, non-viral or viral vectors for suicide gene therapy of cancer make more informed decisions when choosing enzyme/prodrug systems. PMID:25575867
Is there any possibility of detecting the use of genetic engineering in processed foods?
Greiner, R; Konietzny, U; Jany, K D
To elucidate if there is any possibility to identify highly processed foods as produced through genetic engineering, beer, soya bean oil, processed tomato (ketch-up, paste, pizza tomatoes, peeled tomatoes, soup) and potato (french fries, crisps, mashed potatoes, flour, starch, fried potatoes) products as well as an enzyme preparation (Natuphos) were investigated by PCR. In pizza tomatoes, peeled tomatoes, french fries, fried potatoes, potato flour and potato crisps DNA suitable for PCR was found. Therefore, it is possible to identify these products as produced through genetic engineering. Such an identification is impossible in certain beers (pilsener, export, Nutfield lyte), soya bean oil, tomato soup, potato starch, mashed potatoes and Natuphos since PCR-analysis gave no indication of the presence of DNA in these products. As it was shown by adding Escherichia coli DNA the used method is, in principle, capable of detecting specifically small amounts of DNA in such products. PMID:9246732
Bone Marrow Transplantation in Mice as a Tool to Generate Genetically Modified Animals
NASA Astrophysics Data System (ADS)
R?szer, Tamßs; Pintye, ╔va; Benk?, Ilona
Transgenic mice can be used either as models of known inherited human diseases or can be applied to perform phenotypic tests of genes with unknown function. In some special applications of gene modification we have to create a tissue specific mutation of a given gene. In some cases however the gene modification can be lethal in the intrauterine life, therefore we should engraft the mutated cells in the postnatal life period. After total body irradiation transplantation of bone marrow cells can be a solution to introduce mutant hematopoietic stem cells into a mature animal. Bone marrow transplantation is a useful and novel tool to study the role of hematopoietic cells in the pathogenesis of inflammation, autoimmune syndromes and many metabolic alterations coupled recently to leukocyte functions.
Recombineering: highly efficient in vivo genetic engineering using single-strand oligos.
Sawitzke, James A; Thomason, Lynn C; Bubunenko, Mikhail; Li, Xintian; Costantino, Nina; Court, Donald L
Recombineering provides the ability to make rapid, precise, and inexpensive genetic alterations to any DNA sequence, either in the chromosome or cloned onto a vector that replicates in E. coli (or other recombineering-proficient bacteria), and to do so in a highly efficient manner. Complicated genetic constructs that are impossible to make with in vitro genetic engineering can be created in days with recombineering. Recombineering with single-strand DNA (ssDNA) can be used to create single or multiple clustered point mutations, small or large (up to 10kb) deletions, and small (10-20 base) insertions such as sequence tags. Using optimized conditions, point mutations can be made with such high frequencies that they can be found without selection. This technology excels at creating both directed and random mutations. PMID:24182922
Tipping Points in Seaweed Genetic Engineering: Scaling Up Opportunities in the Next Decade
Lin, Hanzhi; Qin, Song
Seaweed genetic engineering is a transgenic expression system with unique features compared with those of heterotrophic prokaryotes and higher plants. This study discusses several newly sequenced seaweed nuclear genomes and the necessity that research on vector design should consider endogenous promoters, codon optimization, and gene copy number. Seaweed viruses and artificial transposons can be applied as transformation methods after acquiring a comprehensive understanding of the mechanism of viral infections in seaweeds and transposon patterns in seaweed genomes. After cultivating transgenic algal cells and tissues in a photobioreactor, a biosafety assessment of genetically modified (GM) seaweeds must be conducted before open-sea application. We propose a set of programs for the evaluation of gene flow from GM seaweeds to local/geographical environments. The effective implementation of such programs requires fundamentally systematic and interdisciplinary studies on algal physiology and genetics, marine hydrology, reproductive biology, and ecology. PMID:24857961
Genetically engineered multivalent single chain antibody constructs for cancer therapy
Surinder Batra, Ph.D.
Current therapeutic approaches against the advanced stages of human solid tumors are palliative rather than curative. Many modalities, including, surgery, radiation, and chemotherapy, either alone or in combination have met with only modest success for advanced metastatic cancers. Radioimmunotherapy (RIT) combines the specificity of monoclonal antibodies with cytotxic effects of radioisotopes. It is the ?smart? way of delivering radiation to the known and occult metastatic cancer cells and is independent of drug toxicity and/or hormone resistance. The tumor associated glycoprotein-72 (TAG-72) containing the unique disaccharide sialyl-Tn, is highly expressed in majority of adenocarcinomas, including carcinomas of the prostate, breast, ovaries, pancreas and colon (80-90%) compared to undetectable expression in normal tissues. Monoclonal antibody CC49, reactive with TAG-72, after conjugation to potent gamma- and beta-emitting radionuclides, has been useful in selective systemic radiolocalization of disease and therapy of primary and metastatic tumor sites. However, limited therapeutic responses were observed in patients. Limited success of antibody based delivery of radioisotopes can be attributed to several factors including undesirable pharmacokinetics, poor tumor uptake and high immunogenicity of intact antibodies (IgGs). The primary factors contributing towards the failure of RIT include: 1) longer serum half-lives of the intact IgG molecules resulting in the radiotoxicity, 2) generation of human antibodies against murine antibodies (HAMA) that limits the frequency of dose administration, 3) poor diffusion rates of intact IgG due to the large size and 4) high interstitial fluid pressures (IFP) encountered in solid tumors. The major goal of our multidisciplinary project was to develop specific novel radiopharmaceuticals, with desired pharmacokinetics, for the diagnosis and therapy of solid tumors. To overcome the low uptake of radioactivity by tumors and to increase its tumor: normal tissue ratio for improved therapeutic index, we engineered a variety antibody constructs. These constructs were evaluated using novel approaches like special radionuclides, pretargeting and optimization. Due to the smaller size, the engineered antibody molecules should penetrate better throughout a tumor mass, with less dose heterogeneity, than is the case with intact IgG. Multivalent scFvs with an appropriate radionuclide, therefore, hold promising prospects for cancer therapy and clinical imaging in MAb-based radiopharmaceuticals. In addition, the human anti-mouse antibodies (HAMA) responses in patients against antibody-based therapy are usually directed against the immunoglobulin constant regions; however, anti-idiotypic responses can also be detected. The HAMA responses reduce the efficacy of treatment by removing the circulating antibody molecules, fragments, and possibly scFvs by altering the pharmacokinetic properties of the antibody. HAMA responses against divalent IgG, divalent Ig fragments, and possibly multimeric scFvs could cause immune complex formation with hypersensitivity or allergic reactions that could be harmful to patients. The use of small molecules, such as scFvs (monomeric as well as multimeric), with their shorter biological half-lives and the lack of the constant regions and humanized variable (binding regions) performed in our studies should reduce the development of HAMA. The generation of humanized and fully human scFvs should further reduce the development of HAMA. Specific accomplishments on the project are the production of large amounts of recombinant antibodies as they are required in large amounts for cancer diagnosis and therapy. A variety of single-chain Fv (scFv) constructs were engineered for the desired pharmacokinetic properties. Tetrameric and dimeric scFvs showed a two-fold advantage: (1) there was a considerable gain in avidity as compared to smaller fragments, and (2) the biological half-life was more compatible with RIT and RIS requirements. For RIT, delivery for sc(Fv)2 and [sc(Fv)2]2 in a fr
Production of polyhydroxyalkanoates from intact triacylglycerols by genetically engineered Pseudomonas.
Solaiman, D K; Ashby, R D; Foglia, T A
Pseudomonas putida and P oleovorans have been extensively studied for their production of medium-chain-length (mcl)-polyhydroxyalkanoates (PHA). These bacteria are incapable of metabolizing triacylglycerols (TAGs). We have constructed recombinant P. putida and P. oleovorans that can utilize TAGs as substrates for growth and mcl-PHA synthesis. A recombinant plasmid, pCN51lip-1, carrying Pseudomonas lipase genes was used to electrotransform these organisms. The transformants expressed TAG-hydrolyzing activity as shown by a rhodamine B fluorescence plate assay. The genetically modified organisms grew in TAG-containing medium to a cell dry weight of 2-4 g/l. The recombinant P. putida produced mcl-PHA at a crude yield of 0.9-1.6 g/l with lard or coconut oil (Co) as substrate. While P. oleovorans transformant did not produce mcl-PHA, a mixed-culture fermentation approach with the wild-type and recombinant strains afforded polymer production from Co at a crude yield of 0.5 g/l. Compositional analysis by gas chromatography/mass spectrometry showed that beta-hydroxyoctanoate (31-45 mol %) and beta-hydroxydecanoate (28-35 mol %) were the dominant repeat units of the TAG-based PHA. The number-average and weight-average molecular masses of the PHAs as determined by gel permeation chromatography were 82-170 x 10(3) g/mol and 464-693 x 10(3) g/mol, respectively. The recombinant approach can greatly increase the number of organisms that can be used to produce PHA from fat and oil substrates. PMID:11601611
Genetic engineering of bacteria and their potential for Hg 2+ bioremediation
Shaolin Chen; David B. Wilson
Ion exchange or biosorptive processes for metalremoval generally lack specificity in metal bindingand are sensitive to ambient conditions, e.g. pH,ionic strength and the presence of metal chelators. Inthis study, cells of a genetically engineered Escherichia coli strain, JM109, which expressesmetallothionein and a Hg2+ transport system afterinduction were evaluated for their selectivity forHg2+ accumulation in the presence of sodium,magnesium, or cadmium
Genetically-Engineered Pig-to-Baboon Liver Xenotransplantation: Histopathology of Xenografts and Native Organs
Burcin Ekser; Edwin Klein; Jing He; Donna B. Stolz; Gabriel J. Echeverri; Cassandra Long; Chih Che Lin; Mohamed Ezzelarab; Hidetaka Hara; Massimiliano Veroux; David Ayares; David K. C. Cooper; Bruno Gridelli
Orthotopic liver transplantation was carried out in baboons using wild-type (WT, n = 1) or genetically-engineered pigs (?1,3-galactosyltransferase gene-knockout, GTKO), n = 1; GTKO pigs transgenic for human CD46, n = 7) and a clinically-acceptable immunosuppressive regimen. Biopsies were obtained from the WT pig liver pre-Tx and at 30 min, 1, 2, 3, 4 and 5 h post-transplantation. Biopsies of
Systemic Delivery of Human Growth Hormone by Injection of Genetically Engineered Myoblasts
Jyotsna Dhawan; Lydia C. Pan; Grace K. Pavlath; Marilyn A. Travis; Andrea M. Lanctot; Helen M. Blau
A recombinant gene encoding human growth hormone (hGH) was stably introduced into cultured myoblasts with a retroviral vector. After injection of genetically engineered myoblasts into mouse muscle, hGH could be detected in serum for 3 months. The fate of injected myoblasts was assessed by coinfecting the cells with two retroviral vectors, one encoding hGH and the other encoding beta-galactosidase from
Continuous delivery of human and mouse erythropoietin in mice by genetically engineered polymer encapsulated myoblasts
E Rķgulier; BL Schneider; N Dķglon; Y Beuzard; P Aebischer
The transplantation of polymer encapsulated myoblasts genetically engineered to secrete erythropoietin (Epo) may obviate the need for repeated parenteral administration of recombinant Epo as a treatment for chronic renal failure, cancer or AIDS-associated anemia. To explore this possibility, the human and mouse Epo cDNAs under the control of the housekeeping mouse PGK-1 promoter were transfected into mouse C2C12 myoblasts, which
366. Suicide Gene Therapy of Glioma Using Genetically Engineered Human Mesenchymal Stem Cells
Hiroki Namba; Shaoyi Li; Tsutomu Tokuyama; Junkoh Yamamoto; Naoki Yokota
We tested anti-tumor activity of genetically engineered mesenchymal stem cells (MSCs) derived from the human adult bone marrow against glioma cells. MSCs were transduced with the herpes simplex virus-thymidine kinase gene (MSCtk cells) and mixed with various glioma cell lines (two human glioma cell lines, A-172 and T98G and C6 rat glioma cell line) and examined in vitro and in
A Comparative Risk Assessment of Genetically Engineered, Mutagenic, and Conventional Wheat Production Systems
Robert K. D. Peterson; Leslie M. Shama
Wheat (Triticum aestivum L.) varieties produced using modern biotechnologies, such as genetic engineering and mutagenic techniques, have lagged behind\\u000a other crop species, but are now being developed and, in the case of mutagenic wheat, commercially grown around the world.\\u000a Because these wheat varieties have emerged recently, there is a unique opportunity to assess comparatively the potential environmental\\u000a risks (human health,
Effects of sugar concentration on recombinant human ? 1-antitrypsin production by genetically engineered rice cell
Masaaki Terashima; Yoko Ejiri; Naohiro Hashikawa; Hiroyuki Yoshida
Productivity of recombinant human ?1-antitrypsin (rAAT) with a genetically engineered rice cell using an inducible promoter has been studied by batch-wise and continuous production. A simple model explained the effect of proteases released from the disrupted cells on the rAAT degradation. Glucose concentration in the medium significantly affected the rAAT productivity in the continuous production, because the rAAT was induced
In vivo cervical cancer growth inhibition by genetically engineered cytotoxic T cells
Peter Dall; Isabell Herrmann; Bettina Durst; Mariam A. Stoff-Khalili; Gerd Bauerschmitz; Bettina Hanstein; Dieter Niederacher
Purpose: The CD44 v7\\/8 splice variant that is frequently expressed in cervical carcinoma and rarely expressed in normal tissues displays promising properties as a target antigen for cancer immune therapy. In this study, cytotoxic T lymphocytes (CTLs) were genetically engineered to gain CD44v7\\/8 target specificity. Methods: Clone 96 (CI96), an established murine cytotoxic T-cell line, and na've murine T cells
Pacific Northwest Laboratory compared intact soil-core microcosms and the field for ecosystem structural and functional properties after the introduction of a model genetically engineered microorganism (GEM). This project used two distinct microbial types as model GEMs, Gram nega...
Reproductive cloning, genetic engineering and the autonomy of the child: the moral agent and the open future.
Mameli, M
Some authors have argued that the human use of reproductive cloning and genetic engineering should be prohibited because these biotechnologies would undermine the autonomy of the resulting child. In this paper, two versions of this view are discussed. According to the first version, the autonomy of cloned and genetically engineered people would be undermined because knowledge of the method by which these people have been conceived would make them unable to assume full responsibility for their actions. According to the second version, these biotechnologies would undermine autonomy by violating these people's right to an open future. There is no evidence to show that people conceived through cloning and genetic engineering would inevitably or even in general be unable to assume responsibility for their actions; there is also no evidence for the claim that cloning and genetic engineering would inevitably or even in general rob the child of the possibility to choose from a sufficiently large array of life plans. PMID:17264194
Intact soil-core microcosms were studied to determine their applicability for evaluating the transport, survival and potential ecosystem effects of genetically engineered microorganisms before they are released into the environment. oi1-core microcosms were planted with wheat and...
Dr. Campbell's Bio111 Exam #4 Spring 2007 Biology 111 Take-Home Exam #4 Cancer, HIV, & Genetic Engineering
Campbell, A. Malcolm
, & Genetic Engineering There is no time limit on this test, though I have tried to design one that you should examples in your answer. 8 pts. 3) List four major types of proto-oncogenes and give a specific human
A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi
N°dvig, Christina S.; Nielsen, Jakob B.; Kogle, Martin E.; Mortensen, Uffe H.
The number of fully sequenced fungal genomes is rapidly increasing. Since genetic tools are poorly developed for most filamentous fungi, it is currently difficult to employ genetic engineering for understanding the biology of these fungi and to fully exploit them industrially. For that reason there is a demand for developing versatile methods that can be used to genetically manipulate non-model filamentous fungi. To facilitate this, we have developed a CRISPR-Cas9 based system adapted for use in filamentous fungi. The system is simple and versatile, as RNA guided mutagenesis can be achieved by transforming a target fungus with a single plasmid. The system currently contains four CRISPR-Cas9 vectors, which are equipped with commonly used fungal markers allowing for selection in a broad range of fungi. Moreover, we have developed a script that allows identification of protospacers that target gene homologs in multiple species to facilitate introduction of common mutations in different filamentous fungi. With these tools we have performed RNA-guided mutagenesis in six species of which one has not previously been genetically engineered. Moreover, for a wild-type Aspergillus aculeatus strain, we have used our CRISPR Cas9 system to generate a strain that contains an AACU_pyrG marker and demonstrated that the resulting strain can be used for iterative gene targeting. PMID:26177455
Towards programming languages for genetic engineering of living cells
Pedersen, Michael; Phillips, Andrew
Synthetic biology aims at producing novel biological systems to carry out some desired and well-defined functions. An ultimate dream is to design these systems at a high level of abstraction using engineering-based tools and programming languages, press a button, and have the design translated to DNA sequences that can be synthesized and put to work in living cells. We introduce such a programming language, which allows logical interactions between potentially undetermined proteins and genes to be expressed in a modular manner. Programs can be translated by a compiler into sequences of standard biological parts, a process that relies on logic programming and prototype databases that contain known biological parts and protein interactions. Programs can also be translated to reactions, allowing simulations to be carried out. While current limitations on available data prevent full use of the language in practical applications, the language can be used to develop formal models of synthetic systems, which are otherwise often presented by informal notations. The language can also serve as a concrete proposal on which future language designs can be discussed, and can help to guide the emerging standard of biological parts which so far has focused on biological, rather than logical, properties of parts. PMID:19369220
Engineered temperature compensation in a synthetic genetic clock
Hussain, Faiza; Gupta, Chinmaya; Hirning, Andrew J.; Ott, William; Matthews, Kathleen S.; Josi?, KreÜimir; Bennett, Matthew R.
Synthetic biology promises to revolutionize biotechnology by providing the means to reengineer and reprogram cellular regulatory mechanisms. However, synthetic gene circuits are often unreliable, as changes to environmental conditions can fundamentally alter a circuitÆs behavior. One way to improve robustness is to use intrinsic properties of transcription factors within the circuit to buffer against intra- and extracellular variability. Here, we describe the design and construction of a synthetic gene oscillator in Escherichia coli that maintains a constant period over a range of temperatures. We started with a previously described synthetic dual-feedback oscillator with a temperature-dependent period. Computational modeling predicted and subsequent experiments confirmed that a single amino acid mutation to the core transcriptional repressor of the circuit results in temperature compensation. Specifically, we used a temperature-sensitive lactose repressor mutant that loses the ability to repress its target promoter at high temperatures. In the oscillator, this thermoinduction of the repressor leads to an increase in period at high temperatures that compensates for the decrease in period due to Arrhenius scaling of the reaction rates. The result is a transcriptional oscillator with a nearly constant period of 48 min for temperatures ranging from 30 ░C to 41 ░C. In contrast, in the absence of the mutation the period of the oscillator drops from 60 to 30 min over the same temperature range. This work demonstrates that synthetic gene circuits can be engineered to be robust to extracellular conditions through protein-level modifications. PMID:24395809
Lipidomic analysis of Arabidopsis seed genetically engineered to contain DHA
Zhou, Xue-Rong; Callahan, Damien L.; Shrestha, Pushkar; Liu, Qing; Petrie, James R.; Singh, Surinder P.
Metabolic engineering of omega-3 long-chain (?C20) polyunsaturated fatty acids (?3 LC-PUFA) in oilseeds has been one of the key targets in recent years. By expressing a transgenic pathway for enhancing the synthesis of the ?3 LC-PUFA docosahexaenoic acid (DHA) from endogenous ?-linolenic acid (ALA), we obtained the production of fish oil-like proportions of DHA in Arabidopsis seed oil. Liquid chromatography-mass spectrometry (LC-MS) was used to characterize the triacylglycerol (TAG), diacylglycerol (DAG) and phospholipid (PL) lipid classes in the transgenic and wild type Arabidopsis seeds at both developing and mature stages. The analysis identified the appearance of several abundant DHA-containing phosphatidylcholine (PC), DAG and TAG molecular species in mature seeds. The relative abundances of PL, DAG, and TAG species showed a preferred combination of LC-PUFA with ALA in the transgenic seeds, where LC-PUFA were esterified in positions usually occupied by 20:1?9. Trace amounts of di-DHA PC and tri-DHA TAG were identified and confirmed by high resolution MS/MS. Studying the lipidome in transgenic seeds provided insights into where DHA accumulated and combined with other fatty acids of neutral and phospholipids from the developing and mature seeds. PMID:25225497
Small-scale field test of the genetically engineered lacZY marker
Hattemer-Frey, H.A.; Brandt, E.J.; Travis, C.C. (Oak Ridge National Laboratory, TN (USA))
Commercial genetic engineering is advancing into areas that require the small-scale introduction of genetically engineered microorganisms (GEMs) to better quantify variables that affect microorganism distribution and survival and to document potential long-term consequences. A recombinant DNA marker system, the lacZY marker, developed by the Monsanto Agricultural Co., enables the distribution and fate of marked fluorescent pseudomonad organisms to be monitored under actual field conditions. Critical evaluation of GEMs under field conditions is imperative if plant-beneficial effects are to be correlated with organism release. This paper evaluates the effectiveness of this marker system and its ability to facilitate the assessment of risks associated with deliberate environmental introductions of genetically engineered microorganisms. Results of prerelease contained growth chamber and field experiments demonstrated that: (1) the scientific risk assessment methodology adopted by Monsanto and approved by the U.S. Environmental Protection Agency was appropriate and comprehensive; (2) the deliberate introduction of a GEM did not pose unacceptable or unforeseen risks to human health or the environment; (3) the lacZY marker is an effective environmental tracking tool; and (4) regulatory oversight should reflect the expected risk and not be excessively burdensome for all GEMs.
Payments for agrobiodiversity conservation services (PACS): Creating incentive mechanisms for the sustained on-farm utilization of plant and animal genetic resources
Ulf Narloch; Unai Pascual; Adam G. Drucker
This paper builds on the experiences from the numerous payment for environmental services (PES) and PES-like schemes in order to establish a framework for evaluating the potential of PES as an incentive mechanism for the conservation of agrobiodiversity per se in poor farming communities. Such - often indigenous - communities conserve much of the world's threatened plant and animal genetic
Genetic and phenotypic evidence of the Salmonella enterica serotype Enteritidis human-animal interface in Chile
Retamal, Patricio; Fresno, Marcela; Dougnac, Catherine; Gutierrez, Sindy; Gornall, Vanessa; Vidal, Roberto; Vernal, Rolando; Pujol, Myriam; Barreto, Marlen; Gonzßlez-Acu±a, Daniel; Abalos, Pedro
Salmonella enterica serotype Enteritidis is a worldwide zoonotic agent that has been recognized as a very important food-borne bacterial pathogen, mainly associated with consumption of poultry products. The aim of this work was to determine genotypic and phenotypic evidence of S. Enteritidis transmission among seabirds, poultry and humans in Chile. Genotyping was performed using PCR-based virulotyping, pulse-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Pathogenicity-associated phenotypes were determined with survival to free radicals, acidic pH, starvation, antimicrobial resistance, and survival within human dendritic cells. As result of PCR and PFGE assays, some isolates from the three hosts showed identical genotypic patterns, and through MLST it was determined that all of them belong to sequence type 11. Phenotypic assays show diversity of bacterial responses among isolates. When results were analyzed according to bacterial host, statistical differences were identified in starvation and dendritic cells survival assays. In addition, isolates from seabirds showed the highest rates of resistance to gentamycin, tetracycline, and ampicillin. Overall, the very close genetic and phenotypic traits shown by isolates from humans, poultry, and seabirds suggest the inter-species transmission of S. Enteritidis bacteria between hosts, likely through anthropogenic environmental contamination that determines infection of seabirds with bacteria that are potentially pathogenic for other susceptible organism, including humans. PMID:26029196
Genetic and phenotypic evidence of the Salmonella enterica serotype Enteritidis human-animal interface in Chile.
Retamal, Patricio; Fresno, Marcela; Dougnac, Catherine; Gutierrez, Sindy; Gornall, Vanessa; Vidal, Roberto; Vernal, Rolando; Pujol, Myriam; Barreto, Marlen; Gonzßlez-Acu±a, Daniel; Abalos, Pedro
Salmonella enterica serotype Enteritidis is a worldwide zoonotic agent that has been recognized as a very important food-borne bacterial pathogen, mainly associated with consumption of poultry products. The aim of this work was to determine genotypic and phenotypic evidence of S. Enteritidis transmission among seabirds, poultry and humans in Chile. Genotyping was performed using PCR-based virulotyping, pulse-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Pathogenicity-associated phenotypes were determined with survival to free radicals, acidic pH, starvation, antimicrobial resistance, and survival within human dendritic cells. As result of PCR and PFGE assays, some isolates from the three hosts showed identical genotypic patterns, and through MLST it was determined that all of them belong to sequence type 11. Phenotypic assays show diversity of bacterial responses among isolates. When results were analyzed according to bacterial host, statistical differences were identified in starvation and dendritic cells survival assays. In addition, isolates from seabirds showed the highest rates of resistance to gentamycin, tetracycline, and ampicillin. Overall, the very close genetic and phenotypic traits shown by isolates from humans, poultry, and seabirds suggest the inter-species transmission of S. Enteritidis bacteria between hosts, likely through anthropogenic environmental contamination that determines infection of seabirds with bacteria that are potentially pathogenic for other susceptible organism, including humans. PMID:26029196
Homozygous; Inheritance; Heterozygous; Inheritance patterns; Heredity and disease; Heritable; Genetic markers ... The chromosomes are made up of strands of genetic information called DNA. Each chromosome contains sections of ...
Genetically engineered polypeptides for inorganics: A utility in biological materials science and engineering
Candan Tamerler; Turgay Kacar; Deniz Sahin; Hanson Fong; Mehmet Sarikaya
Adapting molecular biology to materials science we developed peptide-based protocols for the assembly and formation of hybrid materials and systems. In this approach of generating molecular scale biomimetic materials, peptides are designed, synthesized, genetically tailored and, finally, utilized as potential molecular linkers in self-assembly, ordered organization, and fabrication of inorganics for specific technological applications. The potential areas range from molecular
Which insulin to use? Human or animal?
V. Mohan
The introduction of insulin was a breakthrough in the treatment of diabetes and it produced a remarkable increase in the life expectancy of diabetic patients. Animal-derived insulins have been used to treat pe o- ple with diabetes since insulin was first discovered and continuously subjected to various purification technologies. Genetically engineered human insulin was introduced in 1982 and now the
Environmental concerns associated with the design of genetic engineering facilities.
Watt, J C; Wroniewicz, V S; Ioli, D F
Recombinant DNA technology is being used to produce a wide spectrum of products, such as vaccines, interferon, insulin, and growth hormones. In the design of facilities employing this technology, critical consideration must be given to the protection of the environment, both in the prevention of releases of recombinant DNA organisms into the environment and in the treatment of wastes originating from the production facilities. The design requirements for containment of large-scale systems are complex and require detailed analysis to insure that the containment system can handle both the normal and emergency releases of recombinant DNA organisms. This must include the prevention of releases through either liquid discharges or air emissions. The "killing" method used in the process for either the cells (extracellular product) or the broth (intracellular product) is an important step and can have significant implications in downstream treatment of wastewaters. Since fermentation is the primary process used in the production of recombinant DNA products, wastewater characteristics from this area of the process are basically similar to those of other fermentation processes. They differ, however, because of the "killing" step in the process, which can introduce compounds not normally found in fermentation wastewaters. This can complicate the treatment process by requiring additional treatment operations. Characteristics of wastewaters from other areas of the process can be very diverse, and no general characterization can be made. Techniques for recovery and purification can vary from product to product or even from plant to plant, making characterization difficult. It is important, therefore, that each process be examined in detail so that waste characterization is meaningful and useful in the design of treatment facilities. Because of the complex nature of the processes involved in the production of recombinant DNA products, wastewater treatment can also become a very complex problem. Systems to treat these wastewaters can include many diverse unit operations, from pretreatment of selected streams to tertiary treatment of the combined streams to meet stringent effluent criteria. While biological treatment is almost always applicable, waste loads are very high, and multiple-stage systems could be required. Early and ongoing interface between the process development scientists and engineers and the environmental disciplines allows for the early recognition of potential environmental problems. With early recognition, many of these problems can be economically and efficiently addressed in the design of the facility.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:3178641
Automated, quantitative cognitive/behavioral screening of mice: for genetics, pharmacology, animal cognition and undergraduate instruction.
Gallistel, C R; Balci, Fuat; Freestone, David; Kheifets, Aaron; King, Adam
We describe a high-throughput, high-volume, fully automated, live-in 24/7 behavioral testing system for assessing the effects of genetic and pharmacological manipulations on basic mechanisms of cognition and learning in mice. A standard polypropylene mouse housing tub is connected through an acrylic tube to a standard commercial mouse test box. The test box has 3 hoppers, 2 of which are connected to pellet feeders. All are internally illuminable with an LED and monitored for head entries by infrared (IR) beams. Mice live in the environment, which eliminates handling during screening. They obtain their food during two or more daily feeding periods by performing in operant (instrumental) and Pavlovian (classical) protocols, for which we have written protocol-control software and quasi-real-time data analysis and graphing software. The data analysis and graphing routines are written in a MATLAB-based language created to simplify greatly the analysis of large time-stamped behavioral and physiological event records and to preserve a full data trail from raw data through all intermediate analyses to the published graphs and statistics within a single data structure. The data-analysis code harvests the data several times a day and subjects it to statistical and graphical analyses, which are automatically stored in the "cloud" and on in-lab computers. Thus, the progress of individual mice is visualized and quantified daily. The data-analysis code talks to the protocol-control code, permitting the automated advance from protocol to protocol of individual subjects. The behavioral protocols implemented are matching, autoshaping, timed hopper-switching, risk assessment in timed hopper-switching, impulsivity measurement, and the circadian anticipation of food availability. Open-source protocol-control and data-analysis code makes the addition of new protocols simple. Eight test environments fit in a 48 in x 24 in x 78 in cabinet; two such cabinets (16 environments) may be controlled by one computer. PMID:24637442
Logic Animation and Logic Animation Platform
Mao Chen; Qiang Ge; Qingtang Liu; Zhiguo Si
Logic animation, a new style of animation, was first proposed in the application of dynamic geometry software. The concept, features, and possible application prospects of logic animation are further introduced in details in this paper, compared with the traditional style of animation-time-sequential animation. Based on the design ideas of dynamic geometry software, Flash and game engine, the design of a
Self-association and modification of a genetically engineered polypeptide
NASA Astrophysics Data System (ADS)
Top, Ayben
A genetically synthesized polypeptide and polyethylene glycol (5 kDa or 10 kDa) functionalized forms of its alanine-rich helical domain were characterized. The polypeptide composed of an N-terminal histidine tag, and an alanine-rich domain, denoted as 17H6, has a sequence of: MGH10 SSGHIHM(AAAQEAAAAQAAAQAEAAQAAQ)6AGGYGGMG. 17H6 was originally designed as a scaffold to investigate multivalent interactions after glycosylation through reactive glutamic acid residues. We speculated that the protonation of the glutamic acid residues in these sequences would afford facile opportunities to manipulate their folding and assembly behavior considering the beta-sheet propensities of similar polypeptides at acidic pH. Thus, in the first part of this study, thermal unfolding, reversible self-association, and irreversible aggregation of 17H6 were investigated. Dynamic light scattering, and thermal unfolding measurements indicate that 17H6 spontaneously and reversibly self-associates at an acidic pH and ambient temperature. The resulting multimers have an average hydrodynamic radius of ś 10-20 nm and reversibly dissociate to monomers upon an increase to pH 7.4. Both free monomer and 17H6 chains within the multimers are beta-helical and folded at ambient and sub-ambient temperatures. Reversible unfolding of the monomer occurs upon heating of solutions at pH 7.4. At pH 2.3, heating first causes incomplete dissociation and unfolding of the constituent chains. Further incubation at an elevated temperature (80░C) induces additional structural and morphological changes and results in fibrils with a beta-sheet structure and a width of 5-10 nm (7 nm mean) as observed via transmission electron microscopy (TEM). In the second part, the histidine tag, which imparts solubility to the alanine-rich domain at acidic pH was cleaved. Propionaldehyde-functionalized poly(ethylene glycol) (PEG) molecules (5 kDa or 10 kDa) were attached to the N-terminus of the cleaved polypeptide, c17H6, as a hydrophilic block to compare the effect of these solubility tags on the aggregation and conformation behavior of the alanine-rich domain. Circular dichroic spectroscopy showed that the alpha-helical conformation of the alanine-rich domain was conserved in the conjugates below and near ambient temperatures independent of pH. Similarly, no significant difference between the thermal denaturation behavior of the polypeptide and that of the conjugates was observed at neutral pH. At acidic pH, on the other hand, 17H6 exhibited ś 25% loss in the initial alpha-helical structure upon incubation at 80░C for 3 hours followed by the refolding experiments, whereas the initial alpha-helical content of the conjugates did not change. Comparison of the apparent melting temperatures (ś 54░C for 17H6 and PEG5K-c17H6 and ś 51░C for PEG10K-c17H6) indicated that high temperature stability of the conjugates is not due to the stabilization of the native alpha-helical conformation upon PEGylation. Kinetic experiments at 80 ĪaC for prolonged intervals indicated that PEGylation slowed down the rate of beta-sheet formation and reduced apparent cooperativity. These findings suggest that improved stability of the conjugates at acidic pH is due to the stabilization of the intermediate structures (which is likely to be random coil or early stage of beta-sheet structures) that form prior to the aggregation by reducing the interactions between these intermediates. In contrast to the polypeptide fibrils with ś 7 nm width, TEM images of the conjugates incubated at 80░C for 18 hours showed fibrillar structures with a width of ś 20-30 nm. Thus, it is likely that PEG conjugation also interferes to the arrangement of the polypeptide chains during or prior to the fibril formation. In the third part of this study, the aggregates of the polypeptide and the PEG-polypeptide conjugates that form at the physiological or sub-physiological temperatures and an acidic pH were characterized in detail via dynamic light scattering (DLS), small angle neutron scattering (SANS) and cryogenic transmission electro
Development of a novel genetically modified bioluminescent-bacteria-based assay for detection of fluoroquinolones in animal-derived foods.
Cheng, Guyue; Dong, Xiaobing; Wang, Yulian; Peng, Dapeng; Wang, Xu; Hao, Haihong; Xie, Shuyu; Qu, Wei; Liu, Zhenli; Yuan, Zonghui
Fluoroquinolones (FQNs) are broad-spectrum antibacterial agents widely used in animal husbandry and aquaculture. The residues and antimicrobial resistance of such antibiotics are a major public health concern. To realize multianalyte detection of FQN residues, a genetically modified bacterium, Escherichia coli pK12 harboring plasmid pRecAlux3, was constructed in this study to develop a bioluminescent-bacteria-based assay for the detection of FQNs in animal-derived foods. This assay was based on the principle of induction of an SOS response by FQNs via inducing the recA-promoter-fused luciferase reporter gene existing on the plasmid pRecAlux3. E. coli pK12 was able to recognize 11 FQNs: difloxacin, enrofloxacin, ciprofloxacin, sarafloxacin, norfloxacin, danofloxacin, ofloxacin, pefloxacin, lomefloxacin, marbofloxacin, and orbifloxacin. This method could be applied to 11 edible tissues, including milk, fish muscle, and the muscles, livers, and kidneys of cattle, chickens, and pigs, with a very simple and rapid sample extraction procedure using only phosphate-buffered saline. The limits of detection of the FQNs were between 12.5 and 100Ā?gĀkg(-1), all of which were lower than the maximum residue limits. Most of the recoveries of the FQNs were in the range from 60 to 120Ā%, and the interassay coefficients of variation were less than 30Ā%. This method, confirmed by high-performance liquid chromatography, is reliable and can be used as both a screening test and a semiquantitative assay, when the identity of a single type of FQN is known. PMID:25354889
Engineering modular and tunable genetic amplifiers for scaling transcriptional signals in cascaded gene networks.
Wang, Baojun; Barahona, Mauricio; Buck, Martin
Synthetic biology aims to control and reprogram signal processing pathways within living cells so as to realize repurposed, beneficial applications. Here we report the design and construction of a set of modular and gain-tunable genetic amplifiers in Escherichia coli capable of amplifying a transcriptional signal with wide tunable-gain control in cascaded gene networks. The devices are engineered using orthogonal genetic components (hrpRS, hrpV and PhrpL) from the hrp (hypersensitive response and pathogenicity) gene regulatory network in Pseudomonas syringae. The amplifiers can linearly scale up to 21-fold the transcriptional input with a large output dynamic range, yet not introducing significant time delay or significant noise during signal amplification. The set of genetic amplifiers achieves different gains and input dynamic ranges by varying the expression levels of the underlying ligand-free activator proteins in the device. As their electronic counterparts, these engineered transcriptional amplifiers can act as fundamental building blocks in the design of biological systems by predictably and dynamically modulating transcriptional signal flows to implement advanced intra- and extra-cellular control functions. PMID:25030903
Genetic engineering of crops: a ray of hope for enhanced food security.
Gill, Sarvajeet Singh; Gill, Ritu; Tuteja, Renu; Tuteja, Narendra
Crop improvement has been a basic and essential chase since organized cultivation of crops began thousands of years ago. Abiotic stresses as a whole are regarded as the crucial factors restricting the plant species to reach their full genetic potential to deliver desired productivity. The changing global climatic conditions are making them worse and pointing toward food insecurity. Agriculture biotechnology or genetic engineering has allowed us to look into and understand the complex nature of abiotic stresses and measures to improve the crop productivity under adverse conditions. Various candidate genes have been identified and transformed in model plants as well as agriculturally important crop plants to develop abiotic stress-tolerant plants for crop improvement. The views presented here are an attempt toward realizing the potential of genetic engineering for improving crops to better tolerate abiotic stresses in the era of climate change, which is now essential for global food security. There is great urgency in speeding up crop improvement programs that can use modern biotechnological tools in addition to current breeding practices for providing enhanced food security. PMID:24686131
Genetically engineering cyanobacteria to convert CO?, water, and light into the long-chain hydrocarbon farnesene.
Halfmann, Charles; Gu, Liping; Gibbons, William; Zhou, Ruanbao
Genetically engineered cyanobacteria offer a shortcut to convert CO2 and H2O directly into biofuels and high value chemicals for societal benefits. Farnesene, a long-chained hydrocarbon (C15H24), has many applications in lubricants, cosmetics, fragrances, and biofuels. However, a method for the sustainable, photosynthetic production of farnesene has been lacking. Here, we report the photosynthetic production of farnesene by the filamentous cyanobacterium Anabaena sp. PCC 7120 using only CO2, mineralized water, and light. A codon-optimized farnesene synthase gene was chemically synthesized and then expressed in the cyanobacterium, enabling it to synthesize farnesene through its endogenous non-mevalonate (MEP) pathway. Farnesene excreted from the engineered cyanobacterium volatilized into the flask head space and was recovered by adsorption in a resin column. The maximum photosynthetic productivity of farnesene was 69.1?▒?1.8 ?gĘL(-1)ĘO.D.(-1)Ęd(-1). Compared to the wild type, the farnesene-producing cyanobacterium also exhibited a 60 % higher PSII activity under high light, suggesting increased farnesene productivity in such conditions. We envision genetically engineered cyanobacteria as a bio-solar factory for photosynthetic production of a wide range of biofuels and commodity chemicals. PMID:25301585
A tunable and reversible platform for the intracellular formation of genetically engineered protein microdomains
Pastuszka, Martha K.; Janib, Siti M.; Weitzhandler, Isaac; Okamoto, Curtis T.; Hamm-Alvarez, Sarah; MacKay, J. Andrew
From mitochondria to the nuclear envelope, the controlled assembly of micro and nanostructures is essential for life; however, the level at which we can deliberately engineer the assembly of microstructures within intracellular environments remains primitive. To overcome this obstacle, we present a platform to reversibly assemble genetically-engineered protein microdomains (GEPMs) on the time scale of minutes within living cells. Biologically inspired from the human protein tropoelastin, these protein polymers form a secondary aqueous phase above a tunable transition temperature. This assembly process is easily manipulated to occur at or near physiological temperature by adjusting molecular weight and hydrophobicity. We fused protein polymers to green fluorescent protein (GFP) to visualize their behavior within the cytoplasm. While soluble, these polymers have a similar intracellular diffusion constant as cytosolic proteins at 7.4 ?m2/s; however, above their phase transition temperature, the proteins form distinct microdomains (0.1¢2 ?m) with a reduced diffusion coefficient of 1.1 ?m2/s. Microdomain assembly and disassembly are both rapid processes with half-lives of 3.8 and 1.0 min respectively. Via selection of the protein polymer, the assembly temperature is tunable between 20 and 40 ░C. This approach may be useful to control intracellular formation of genetically engineered proteins and protein complexes into concentrated microdomains. PMID:23088632
Application of TILLING and EcoTILLING as Reverse Genetic Approaches to Elucidate the Function of Genes in Plants and Animals
Barkley, N.A; Wang, M.L
With the fairly recent advent of inexpensive, rapid sequencing technologies that continue to improve sequencing efficiency and accuracy, many species of animals, plants, and microbes have annotated genomic information publicly available. The focus on genomics has thus been shifting from the collection of whole sequenced genomes to the study of functional genomics. Reverse genetic approaches have been used for many years to advance from sequence data to the resulting phenotype in an effort to deduce the function of a gene in the species of interest. Many of the currently used approaches (RNAi, gene knockout, site-directed mutagenesis, transposon tagging) rely on the creation of transgenic material, the development of which is not always feasible for many plant or animal species. TILLING is a non-transgenic reverse genetics approach that is applicable to all animal and plant species which can be mutagenized, regardless of its mating / pollinating system, ploidy level, or genome size. This approach requires prior DNA sequence information and takes advantage of a mismatch endonuclease to locate and detect induced mutations. Ultimately, it can provide an allelic series of silent, missense, nonsense, and splice site mutations to examine the effect of various mutations in a gene. TILLING has proven to be a practical, efficient, and an effective approach for functional genomic studies in numerous plant and animal species. EcoTILLING, which is a variant of TILLING, examines natural genetic variation in populations and has been successfully utilized in animals and plants to discover SNPs including rare ones. In this review, TILLING and EcoTILLING techniques, beneficial applications and limitations from plant and animal studies are discussed. PMID:19452039
Natural plant genetic engineer Agrobacterium rhizogenes: role of T-DNA in plant secondary metabolism.
Chandra, Sheela
Agrobacterium rhizogenes is a natural plant genetic engineer. It is a gram-negative soil bacterium that induces hairy root formation. Success has been obtained in exploring the molecular mechanisms of transferred DNA (T-DNA) transfer, interaction with host plant proteins, plant defense signaling and integration to plant genome for successful plant genetic transformation. T-DNA and corresponding expression of rol genes alter morphology and plant host secondary metabolism. During transformation, there is a differential loss of a few T-DNA genes. Loss of a few ORFs drastically affect the growth and morphological patterns of hairy roots, expression pattern of biosynthetic pathway genes and accumulation of specific secondary metabolites. PMID:22048847
Genetic Engineering of Trypanosoma (Dutonella) vivax and In Vitro Differentiation under Axenic Conditions
D'Archivio, Simon; Medina, Mathieu; Cosson, Alain; Chamond, Nathalie; Rotureau, Brice; Minoprio, Paola; Goyard, Sophie
Trypanosoma vivax is one of the most common parasites responsible for animal trypanosomosis, and although this disease is widespread in Africa and Latin America, very few studies have been conducted on the parasite's biology. This is in part due to the fact that no reproducible experimental methods had been developed to maintain the different evolutive forms of this trypanosome under laboratory conditions. Appropriate protocols were developed in the 1990s for the axenic maintenance of three major animal Trypanosoma species: T. b. brucei, T. congolense and T. vivax. These pioneer studies rapidly led to the successful genetic manipulation of T. b. brucei and T. congolense. Advances were made in the understanding of these parasites' biology and virulence, and new drug targets were identified. By contrast, challenging in vitro conditions have been developed for T. vivax in the past, and this per se has contributed to defer both its genetic manipulation and subsequent gene function studies. Here we report on the optimization of non-infective T. vivax epimastigote axenic cultures and on the process of parasite in vitro differentiation into metacyclic infective forms. We have also constructed the first T. vivax specific expression vector that drives constitutive expression of the luciferase reporter gene. This vector was then used to establish and optimize epimastigote transfection. We then developed highly reproducible conditions that can be used to obtain and select stably transfected mutants that continue metacyclogenesis and are infectious in immunocompetent rodents. PMID:22216367
Skeletal tissue engineeringŚfrom in vitro studies to large animal models
Pieter Buma; Willem Schreurs; Nico Verdonschot
Bone is a tissue with a strong regenerative potential. New strategies for tissue engineering of bone should therefore only focus on defects with a certain size that will not heal spontaneously. In the development of tissue-engineered constructs many variables may play a role, e.g. the source of the cells used, the design and mechanical properties of the scaffold and the
A comparative risk assessment of genetically engineered, mutagenic, and conventional wheat production systems.
Peterson, Robert K D; Shama, Leslie M
Wheat (Triticum aestivum L.) varieties produced using modern biotechnologies, such as genetic engineering and mutagenic techniques, have lagged behind other crop species, but are now being developed and, in the case of mutagenic wheat, commercially grown around the world. Because these wheat varieties have emerged recently, there is a unique opportunity to assess comparatively the potential environmental risks (human health, ecological, and livestock risks) associated with genetically engineered, mutagenic, and conventional wheat production systems. Replacement of traditional herbicides with glyphosate in a glyphosate-tolerant (genetically engineered) wheat system or imazamox in an imidazolinone-tolerant (mutagenic) wheat system may alter environmental risks associated with weed management. Additionally, because both systems rely on plants that express novel proteins, the proteins and plants themselves may impose risks. The purpose of our study was to examine comparatively the multiple aspects of risk associated with different wheat production systems in the US and Canada using the risk assessment paradigm. Specifically, we used tier 1 quantitative and qualitative risk assessment methods to compare specific environmental risks associated with the different wheat production systems. Both glyphosate and imazamox present lower human health and ecological risks than many other herbicides associated with conventional wheat production systems evaluated in this study. The differences in risks were most pronounced when comparing glyphosate and imazamox to herbicides currently with substantial market share. Current weight-of-evidence suggests that the transgenic CP4 EPSPS protein present in glyphosate-tolerant wheat poses negligible risk to humans, livestock, and wildlife. Risk for mutated AHAS protein in imidazolinone-tolerant wheat most likely would be low, but there are not sufficient effect and exposure data to adequately characterize risk. Environmental risks for herbicides were more amenable to quantitative assessments than for the transgenic CP4 EPSPS protein and the mutated AHAS protein. PMID:16315092
Evaluation of terrestrial microcosms for detection, fate, and survival analysis of genetically engineered microorganisms and their recombinant genetic material
Fredrickson, J.K.; Seidler, R.J.
The research included in this document represents the current scientific information available regarding the applicability of terrestrial microcosms and related methodologies for evaluating detection methods and the fate and survival of microorganisms in the environment. The three terrestrial microcosms described in this document were used to evaluate the survival and fate of recombinant bacteria in soils and in association with plant surfaces and insects and their transport through soil with percolating water and root systems, and to test new methods and procedures to improve detection and enumeration of bacteria in soil. Simple (potting soil composed of peat mix and perlite, lacking environmental control and monitoring) and complex microcosms (agricultural soil with partial control and monitoring of environmental conditions) were demonstrated to be useful tools for preliminary assessments of microbial viability in terrestrial ecosystems. These studies evaluated the survival patterns of Enterobacter cloacae (pBR322) in soil and on plant surfaces and the ingestion of this same microorganism by cutworms and survival in the foregut and frass. The Versacore microcosm design was used to monitor the fate and competitiveness of genetically engineered bacteria in soil. Both selective media and gene probes were used successfully to follow the fate of two recombinant Pseudomonas sp. introduced into Versacore microcosms. Intact soil-core microcosms were employed to evaluate the fate and transport of genetically altered Azospirillum sp. and Pseudomonas sp. in soil and the plant rhizosphere. The usefulness of these various microcosms as a tool for risk assessment is underscored by the ease in obtaining soil from a proposed field release site to evaluate subsequent GEM fate and survival.
Traditional Animation Keyframe Animation
Treuille, Adrien
#12;Traditional Animation: The Process Ę Story board Ł Sequence of drawings with descriptions Ł Story board Ł Animatic Ł Final Animation #12;Traditional Animation: The Process Ę Key Frames Ł Draw a fewAnimation Traditional Animation Keyframe Animation Interpolating Rotation Forward
Paris-Sud XI, Universitķ de
A GENETIC BASE FOR ESTIMATING THE GENETIC TRANSMITTING ABILITY OF DAIRY BULLS IN POPULATIONS UNDERGOING GENETIC CHANGE (1) F. N. DICKINSON Acting Leader, Animal Impvovement Pvogvams Laboratory, Animal Physiology and Genetics Institute, Agvicultuval Research Service, United States Department of Agvicultuve
Crystals of Serum Albumin for Use in Genetic Engineering and Rational Drug Design
NASA Technical Reports Server (NTRS)
Carter, Daniel C. (Inventor)
Serum albumin crystal forms have been produced which exhibit superior x-ray diffraction quality. The crystals are produced from both recombinant and wild-type human serum albumin, canine, and baboon serum albumin and allow the performance of drug-binding studies as well as genetic engineering studies. The crystals are grown from solutions of polyethylene glycol or ammonium sulphate within prescribed limits during growth times from one to several weeks and include the following space groups: P2(sub 1), C2, P1.
Mitogen-activated protein kinase-regulated AZI1 ¢ an attractive candidate for genetic engineering
Pitzschke, Andrea; Datta, Sneha; Persak, Helene
Mitogen-activated protein kinases and their targets have been in the limelight of plant stress research. Signaling pathways mediating the responses to multiple stresses deserve particular attention. In a recent study, we reported AZI1, a member of the lipid transfer protein family, to play a role in MPK3-mediated responses to salt stress in Arabidopsis thaliana. MPK3 controls AZI1 at the transcriptional and posttranslational level. The AZI1 protein has several properties that make it very attractive for genetic engineering. A model of multi-level control of AZI1 by MPK3 is proposed, and strategies toward optimizing AZI1 protein properties are briefly discussed. PMID:24518841
The genetics of murine Hox loci: TAMERE, STRING, and PANTHERE to engineer chromosome variants.
Tschopp, Patrick; Duboule, Denis
Following their duplications at the base of the vertebrate clade, Hox gene clusters underwent remarkable sub- and neo-functionalization events. Many of these evolutionary innovations can be associated with changes in the transcriptional regulation of their genes, where an intricate relationship between the structure of the gene cluster and the architecture of the surrounding genomic landscape is at play. Here, we report on a portfolio of in vivo genome engineering strategies in mice, which have been used to probe and decipher the genetic and molecular underpinnings of the complex regulatory mechanisms implemented at these loci. PMID:25151159
Independent Neuronal Origin of Seizures and Behavioral Comorbidities in an Animal Model of a Severe Childhood Genetic Epileptic Encephalopathy
Asinof, Samuel K.; Sukoff Rizzo, Stacey J.; Buckley, Alexandra R.; Beyer, Barbara J.; Letts, Verity A.; Frankel, Wayne N.; Boumil, Rebecca M.
The childhood epileptic encephalopathies (EEÆs) are seizure disorders that broadly impact development including cognitive, sensory and motor progress with severe consequences and comorbidities. Recently, mutations in DNM1 (dynamin 1) have been implicated in two EE syndromes, Lennox-Gastaut Syndrome and Infantile Spasms. Dnm1 encodes dynamin 1, a large multimeric GTPase necessary for activity-dependent membrane recycling in neurons, including synaptic vesicle endocytosis. Dnm1Ftfl or ōfitfulö mice carry a spontaneous mutation in the mouse ortholog of DNM1 and recapitulate many of the disease features associated with human DNM1 patients, providing a relevant disease model of human EEÆs. In order to examine the cellular etiology of seizures and behavioral and neurological comorbidities, we engineered a conditional Dnm1Ftfl mouse model of DNM1 EE. Observations of Dnm1Ftfl/flox mice in combination with various neuronal subpopulation specific cre strains demonstrate unique seizure phenotypes and clear separation of major neurobehavioral comorbidities from severe seizures associated with the germline model. This demonstration of pleiotropy suggests that treating seizures per se may not prevent severe comorbidity observed in EE associated with dynamin-1 mutations, and is likely to have implications for other genetic forms of EE. PMID:26125563
Improving UV resistance and virulence of Beauveria bassiana by genetic engineering with an exogenous tyrosinase gene.
Shang, Yanfang; Duan, Zhibing; Huang, Wei; Gao, Qiang; Wang, Chengshu
Insect pathogenic fungi like Beauveria bassiana have been developed as environmentally friendly biocontrol agents against arthropod pests. However, restrictive environmental factors, including solar ultraviolet (UV) radiation frequently lead to inconsistent field performance. To improve resistance to UV damage, we used Agrobacterium-mediated transformation to engineer B. bassiana with an exogenous tyrosinase gene. The results showed that the mitotically stable transformants produced larger amounts of yellowish pigments than the wild-type strain, and these imparted significantly increased UV-resistance. The virulence of the transgenic isolate was also significantly increased against the silkworm Bombyx mori and the mealworm Tenebrio molitor. This study demonstrated that genetic engineering of B. bassiana with a tyrosinase gene is an effective way to improve fungal tolerance against UV damage. PMID:22024554
Milestones in chloroplast genetic engineering: an environmentally friendly era in biotechnology
Daniell, Henry; Khan, Muhammad S.; Allison, Lori
Chloroplast genomes defied the laws of Mendelian inheritance at the dawn of plant genetics, and continue to defy the mainstream approach to biotechnology, leading the field in an environmentally friendly direction. Recent success in engineering the chloroplast genome for resistance to herbicides, insects, disease and drought, and for production of biopharmaceuticals, has opened the door to a new era in biotechnology. The successful engineering of tomato chromoplasts for high-level transgene expression in fruits, coupled to hyper-expression of vaccine antigens, and the use of plant-derived antibiotic-free selectable markers, augur well for oral delivery of edible vaccines and biopharmaceuticals that are currently beyond the reach of those who need them most. PMID:11832280
Formation mechanism of chalcogenide nanocrystals confined inside genetically engineered virus-like particles
Zhou, Ziyou; Bedwell, Gregory J.; Li, Rui; Prevelige, Peter E.; Gupta, Arunava
Engineered virus-like particles (VLP) are attractive for fabricating nanostructured materials for applications in diverse areas such as catalysis, drug delivery, biomedicine, composites, etc. Basic understanding of the interaction between the inorganic guest and biomolecular host is thus important for the controlled synthesis of inorganic nanoparticles inside VLP and rational assembly of ordered VLP-based hierarchical nanostructures. We have investigated in detail the formation mechanism and growth kinetics of semiconducting nanocrystals confined inside genetically engineered bacteriophage P22 VLP using semiconducting CdS as a prototypical example. The selective nucleation and growth of CdS at the engineered sites is found to be uniform during the early stage, followed by a more stochastic growth process. Furthermore, kinetic studies reveal that the presence of an engineered biotemplate helps in significantly retarding the reaction rate. These findings provide guidance for the controlled synthesis of a wide range of other inorganic materials confined inside VLP, and are of practical importance for the rational design of VLP-based hierarchical nanostuctures. PMID:24452221
Targeted genetic manipulations of neuronal subtypes using promoter-specific combinatorial AAVs in wild-type animals
Gompf, Heinrich S.; Budygin, Evgeny A.; Fuller, Patrick M.; Bass, Caroline E.
Techniques to genetically manipulate the activity of defined neuronal subpopulations have been useful in elucidating function, however applicability to translational research beyond transgenic mice is limited. Subtype targeted transgene expression can be achieved using specific promoters, but often currently available promoters are either too large to package into many vectors, in particular adeno-associated virus (AAV), or do not drive expression at levels sufficient to alter behavior. To permit neuron subtype specific gene expression in wildtype animals, we developed a combinatorial AAV targeting system that drives, in combination, subtype specific Cre-recombinase expression with a strong but non-specific Cre-conditional transgene. Using this system we demonstrate that the tyrosine hydroxylase promoter (TH-Cre-AAV) restricted expression of channelrhodopsin-2 (EF1?-DIO-ChR2-EYFP-AAV) to the rat ventral tegmental area (VTA), or an activating DREADD (hSyn-DIO-hM3Dq-mCherry-AAV) to? the? rat? locus? coeruleus? (LC). High expression levels were achieved in both regions. Immunohistochemistry (IHC) showed the majority of ChR2+ neurons (>93%) colocalized with TH in the VTA, and optical stimulation evoked striatal dopamine release. Activation of TH neurons in the LC produced sustained EEG and behavioral arousal. TH-specific hM3Dq expression in the LC was further compared with: (1) a Cre construct driven by a strong but non-specific promoter (non-targeting); and (2) a retrogradely-transported WGA-Cre delivery mechanism (targeting a specific projection). IHC revealed that the area of c-fos activation after CNO treatment in the LC and peri-LC neurons appeared proportional to the resulting increase in wakefulness (non-targeted > targeted > ACC to LC projection restricted). Our dual AAV targeting system effectively overcomes the large size and weak activity barrier prevalent with many subtype specific promoters by functionally separating subtype specificity from promoter strength.
Vision system for animal cell recognition in a bio-engineering process
Toshio FUKUDA; Mitsuru ISHIZUKA; Osamu HASEGAWA; Hajime ASAMA; T. Nagumune; I. Endo
An expert system for the recognition and counting of cells using image processing for mass production of interferon beta is described. The biological cells which make interferon beta are taken from animals and cultured on microcarriers, plastic balls\\/beads 150-200 ?m in diameter floating in a culture solution. Cell images are obtained through a microscope connected to a TV camera. The
STATISTICAL ANALYSIS OF CROSSBREEDING EXPERIMENTS IN PIGS E. BRUNS. -Institut of Animal Breeding and Genetics Gttingen, BRD.
Paris-Sud XI, Universitķ de
STATISTICAL ANALYSIS OF CROSSBREEDING EXPERIMENTS IN PIGS E. BRUNS. - Institut of Animal Breeding CROSSBREEDING PARAMETERS WHEN TWO BREEDS UTILIZE COMMON BREEDING ANIMALS E. FIMLAND. Department of Animal is more expensive in breeds with high rates of difficult calving than in those with low rates. ESTIMATING
STATISTICAL ANALYSIS OF CROSSBREEDING EXPERIMENTS IN PIGS E. BRUNS. -Institut of Animal Breeding and Genetics Gttingen, BRD.
Paris-Sud XI, Universitķ de
STATISTICAL ANALYSIS OF CROSSBREEDING EXPERIMENTS IN PIGS E. BRUNS. - Institut of Animal Breeding is more expensive in breeds with high rates of difficult calving than in those with low rates. ESTIMATING CROSSBREEDING PARAMETERS WHEN TWO BREEDS UTILIZE COMMON BREEDING ANIMALS E. FIMLAND. Department of Animal
Engineering modular and orthogonal genetic logic gates for robust digital-like synthetic biology.
Wang, Baojun; Kitney, Richard I; Joly, Nicolas; Buck, Martin
Modular and orthogonal genetic logic gates are essential for building robust biologically based digital devices to customize cell signalling in synthetic biology. Here we constructed an orthogonal AND gate in Escherichia coli using a novel hetero-regulation module from Pseudomonas syringae. The device comprises two co-activating genes hrpR and hrpS controlled by separate promoter inputs, and a ?(54)-dependent hrpL promoter driving the output. The hrpL promoter is activated only when both genes are expressed, generating digital-like AND integration behaviour. The AND gate is demonstrated to be modular by applying new regulated promoters to the inputs, and connecting the output to a NOT gate module to produce a combinatorial NAND gate. The circuits were assembled using a parts-based engineering approach of quantitative characterization, modelling, followed by construction and testing. The results show that new genetic logic devices can be engineered predictably from novel native orthogonal biological control elements using quantitatively in-context characterized parts. PMID:22009040
Induction of erythropoiesis using human vascular networks genetically engineered for controlled erythropoietin release
Lin, Ruei-Zeng; Dreyzin, Alexandra; Aamodt, Kristie; Li, Dan; Jaminet, Shou-Ching S.; Dudley, Andrew C.
For decades, autologous ex vivo gene therapy has been postulated as a potential alternative to parenteral administration of recombinant proteins. However, achieving effective cellular engraftment of previously retrieved patient cells is challenging. Recently, our ability to engineer vasculature in vivo has allowed for the introduction of instructions into tissues by genetically modifying the vascular cells that build these blood vessels. In the present study, we genetically engineered human blood¢derived endothelial colony-forming cells (ECFCs) to express erythropoietin (EPO) under the control of a tetracycline-regulated system, and generated subcutaneous vascular networks capable of systemic EPO release in immunodeficient mice. These ECFC-lined vascular networks formed functional anastomoses with the mouse vasculature, allowing direct delivery of recombinant human EPO into the bloodstream. After activation of EPO expression, erythropoiesis was induced in both normal and anemic mice, a process that was completely reversible. This approach could relieve patients from frequent EPO injections, reducing the medical costs associated with the management of anemia. We propose this ECFC-based gene-delivery strategy as a viable alternative technology when routine administration of recombinant proteins is needed. PMID:21937702
Induction of erythropoiesis using human vascular networks genetically engineered for controlled erythropoietin release.
Lin, Ruei-Zeng; Dreyzin, Alexandra; Aamodt, Kristie; Li, Dan; Jaminet, Shou-Ching S; Dudley, Andrew C; Melero-Martin, Juan M
For decades, autologous ex vivo gene therapy has been postulated as a potential alternative to parenteral administration of recombinant proteins. However, achieving effective cellular engraftment of previously retrieved patient cells is challenging. Recently, our ability to engineer vasculature in vivo has allowed for the introduction of instructions into tissues by genetically modifying the vascular cells that build these blood vessels. In the present study, we genetically engineered human blood-derived endothelial colony-forming cells (ECFCs) to express erythropoietin (EPO) under the control of a tetracycline-regulated system, and generated subcutaneous vascular networks capable of systemic EPO release in immunodeficient mice. These ECFC-lined vascular networks formed functional anastomoses with the mouse vasculature, allowing direct delivery of recombinant human EPO into the bloodstream. After activation of EPO expression, erythropoiesis was induced in both normal and anemic mice, a process that was completely reversible. This approach could relieve patients from frequent EPO injections, reducing the medical costs associated with the management of anemia. We propose this ECFC-based gene-delivery strategy as a viable alternative technology when routine administration of recombinant proteins is needed. PMID:21937702
Two Concepts of Dignity for Humans and Non-Human Organisms in the Context of Genetic Engineering
Philipp Balzer; Klaus Peter Rippe; Peter Schaber
The 1992 incorporation of an article by referendum in the SwissConstitution mandating that the federal government issue regulations onthe use of genetic material that take into account the dignity ofnonhuman organism raises philosophical questions about how we shouldunderstand what is meant by ``the dignity of nonhuman animals,'' andabout what sort of moral demands arise from recognizing this dignitywith respect to
Space mutagenesis of genetically engineered bacteria expressing recombinant human interferon ?1b and screening of higher yielding strains.
Wang, Junfeng; Liu, Changting; Liu, Jinyi; Fang, Xiangqun; Xu, Chen; Guo, Yinghua; Chang, De; Su, Longxiang
The aim of this study was to investigate the space mutagenesis of genetically engineered bacteria expressing recombinant human interferon ?1b. The genetically engineered bacteria expressing the recombinant interferon ?1b were sent into outer space on the Chinese Shenzhou VIII spacecraft. After the 17 day space flight, mutant strains that highly expressed the target gene were identified. After a series of screening of spaceflight-treated bacteria and the quantitative comparison of the mutant strains and original strain, we found five strains that showed a significantly higher production of target proteins, compared with the original strain. Our results support the notion that the outer space environment has unique effects on the mutation breeding of microorganisms, including genetically engineered strains. Mutant strains that highly express the target protein could be obtained through spaceflight-induced mutagenesis. PMID:24096450
The impact of genetic modification of human foods in the 21st century
Stella G. Uzogara
Genetic engineering of food is the science which involves deliberate modification of the genetic material of plants or animals. It is an old agricultural practice carried on by farmers since early historical times, but recently it has been improved by technology. Many foods consumed today are either genetically modified (GM) whole foods, or contain ingredients derived from gene modification technology.
Practical animal breeding as the key to an integrated view of genetics, eugenics and evolutionary theory: Arend L. Hagedoorn (1885-1953).
Theunissen, Bert
In the history of genetics Arend Hagedoorn (1885-1953) is mainly known for the 'Hagedoorn effect', which states that part of the changes in variability that populations undergo over time are due to chance effects. Leaving this contribution aside, Hagedoorn's work has received scarcely any attention from historians. This is mainly due to the fact that Hagedoorn was an expert in animal breeding, a field that historians have only recently begun to explore. His work provides an example of how a prominent geneticist envisaged animal breeding to be reformed by the new science of heredity. Hagedoorn, a pupil of Hugo de Vries, tried to integrate his insights as a Mendelian geneticist and an animal breeding expert in a unified view of heredity, eugenics and evolution. In this paper I aim to elucidate how these fields were connected in Hagedoorn's work. PMID:24747808
Patterning and transferring hydrogel-encapsulated bacterial cells for quantitative analysis of synthetically engineered genetic circuits.
Choi, Woon Sun; Kim, Minseok; Park, Seongyong; Lee, Sung Kuk; Kim, Taesung
We describe a hydrogel patterning and transferring (HPT) method that facilitates the quantitative analysis of synthetically engineered genetic circuits within bacterial cells. The HPT method encapsulates cells in the alginate hydrogel patterns by using polydimethylsiloxane (PDMS) template. Then, the hydrogel-encapsulated cell patterns are transferred onto an agarose hydrogel substrate that encapsulates inducer chemicals or bacterial cells. Using the HPT method, we demonstrate that inducers in the agarose hydrogel substrate regulate gene expression of the patterned cells for qualitative analysis by activating the promoters of fluorescence protein genes. In addition, we demonstrate that the HPT method can be used for the analysis of the cross-talk between genetic circuits and the concentration-dependent gene expression and regulation because the agarose hydrogel substrate can produce concentration gradients of inducers. Lastly, we demonstrate that the HPT method can be applied to investigating intercellular communication between neighboring cells with a wide range of cell densities. Since the HPT method is simple to deal with but versatile and powerful to quantitatively analyze genetic circuits in living cells in many controllable manners, we believe that the method can be widely used for the rapid advancement of synthetic, molecular, and systems biology. PMID:22014463
Communication support for systems engineering - process modelling and animation with APRIL
Stephan Philippi; Hermann Josef Hill
The most important task in the early stages of systems engineering is the building of models which capture the relevant knowledge of a given application domain. A working communication with domain experts who possess this knowledge is crucial, since misunderstandings almost always lead to expensive system redesigns in later development stages. In this context, especially the modelling of systems behaviour
Evaluating Learning and Attitudes on Tissue Engineering: A Study of Children Viewing Animated Digital Dome Shows Detailing the Biomedicine of Tissue Engineering
Wilson, Anna C.; Gonzalez, Laura L.
Informal science education creates opportunities for the general public to learn about complex health and science topics. Tissue engineering is a fast-growing field of medical science that combines advanced chemistries to create synthetic scaffolds, stem cells, and growth factors that individually or in combination can support the bodies own healing powers to remedy a range of maladies. Health literacy about this topic is increasingly important as our population ages and as treatments become more technologically advanced. We are using a science center planetarium as a projection space to engage and educate the public about the science and biomedical research that supports tissue engineering. The purpose of this study was to test the effectiveness of the films that we have produced for part of the science center planetarium demographic, specifically children ranging in age from 7 to 16 years. A two-group pre- and post-test design was used to compare children's learning and attitude changes in response to the two versions of the film. One version uses traditional voice-over narration; the other version uses dialog between two animated characters. The results of this study indicate that children demonstrated increases in knowledge of the topic with either film format, but preferred the animated character version. The percentage change in children's scores on the knowledge questions given before and after viewing the show exhibited an improvement from 23% correct to 61% correct on average. In addition, many of the things that the children reported liking were part of the design process of the art¢science collaboration. Other results indicated that before viewing the shows 77% of the children had not even heard about tissue engineering and only 17% indicated that they were very interested in it, whereas after viewing the shows, 95% indicated that tissue engineering was a good idea. We also find that after viewing the show, 71% of the children reported that the show made them think, 75% enjoyed it, and 89% felt that they learned something. We discuss the potential impact the films might have on public knowledge, health literacy, and attitudes toward the science of tissue engineering. PMID:21943030
A Pseudomonas putida Strain Genetically Engineered for 1,2,3-Trichloropropane Bioremediation
Samin, Ghufrana; Pavlova, Martina; Arif, M. Irfan; Postema, Christiaan P.; Damborsky, Jiri
1,2,3-Trichloropropane (TCP) is a toxic compound that is recalcitrant to biodegradation in the environment. Attempts to isolate TCP-degrading organisms using enrichment cultivation have failed. A potential biodegradation pathway starts with hydrolytic dehalogenation to 2,3-dichloro-1-propanol (DCP), followed by oxidative metabolism. To obtain a practically applicable TCP-degrading organism, we introduced an engineered haloalkane dehalogenase with improved TCP degradation activity into the DCP-degrading bacterium Pseudomonas putida MC4. For this purpose, the dehalogenase gene (dhaA31) was cloned behind the constitutive dhlA promoter and was introduced into the genome of strain MC4 using a transposon delivery system. The transposon-located antibiotic resistance marker was subsequently removed using a resolvase step. Growth of the resulting engineered bacterium, P. putida MC4-5222, on TCP was indeed observed, and all organic chlorine was released as chloride. A packed-bed reactor with immobilized cells of strain MC4-5222 degraded >95% of influent TCP (0.33 mM) under continuous-flow conditions, with stoichiometric release of inorganic chloride. The results demonstrate the successful use of a laboratory-evolved dehalogenase and genetic engineering to produce an effective, plasmid-free, and stable whole-cell biocatalyst for the aerobic bioremediation of a recalcitrant chlorinated hydrocarbon. PMID:24973068
Correlations for genetic expression for growth of calves of Hereford and Angus dams using a multivariate animal model.
N·±ez-Dominguez, R; Van Vleck, L D; Boldman, K G; Cundiff, L V
The purpose of this study was to estimate the correlation between the expression of genes from sires in purebred and crossbred progeny (rPC) and in Hereford and Angus F1 calves (rHA). Performance traits were weights at birth, 200 d, and 365 d. Progeny from Hereford, Polled Hereford, and Angus bulls mated to Hereford or Angus cows were used to estimate rPC. Progeny from Charolais, Shorthorn, Simmental, Limousin, Maine-Anjou, Chianina, Gelbvieh, Tarentaise, and Salers bulls mated to Hereford or Angus cows were used to estimate rHA. Performances in purebreds (P) and crosses (C) or in Hereford (H) and Angus (A) F1 calves were treated as separate traits. A multivariate animal model with birth year-cow age-sex subclasses, random correlated direct and maternal additive genetic effects, and maternal permanent environmental effects was used. Separate analyses were done by breed of sire. A derivative-free algorithm was used to obtain REML estimates of (co)variance components. Weighted averages across breeds of estimates of heritability for P, C, H, and A were, respectively, .61, .51, .47, and .40 for birth weight, .41, .46, .37, and .34 for weaning weight, and .50, .49, .42, and .46 for yearling weight. Estimates of rPC ranged from .88 to .97, .55 to .94, and .68 to .86 for weights at birth, 200 d, and 365 d, respectively. Estimates of rHA ranged from .43 to .99, .56 to .95, and .50 to .98 for weights at birth, 200 d, and 365 d, respectively. Weighted averages of estimates of rPC and rHA across sire breeds were, respectively, .93 and .85 for birth weight, .77 and .73 for weaning weight, and .76 and .86 for yearling weight. These results indicate that ranking of sires producing purebreds or crosses, or crossbred calves from different breeds of dams, is approximately the same for birth and yearling weights, but some reranking might occur for weaning weight. PMID:8407645
Animal Watching: Outdoors and In.
ERIC Educational Resources Information Center
McLure, John W.
Describes using domesticated, wild, or feral animals to teach students about nature and animal behavior. Connections can be made with psychology, economics, genetics, history, art, and other disciplines. The study of animal behavior provides opportunities for harmless student experimentation. (SAH)
Genetic mechanisms of antimicrobial resistance identified in Salmonella enterica, Escherichia coli, and Enteroccocus spp. isolated from U.S. food animals
Frye, Jonathan G.; Jackson, Charlene R.
The prevalence of antimicrobial resistance (AR) in bacteria isolated from U.S. food animals has increased over the last several decades as have concerns of AR foodborne zoonotic human infections. Resistance mechanisms identified in U.S. animal isolates of Salmonella enterica included resistance to aminoglycosides (e.g., alleles of aacC, aadA, aadB, ant, aphA, and StrAB), ?-lactams (e.g., blaCMY?2, TEM?1, PSE?1), chloramphenicol (e.g., floR, cmlA, cat1, cat2), folate pathway inhibitors (e.g., alleles of sul and dfr), and tetracycline [e.g., alleles of tet(A), (B), (C), (D), (G), and tetR]. In the U.S., multi-drug resistance (MDR) mechanisms in Salmonella animal isolates were associated with integrons, or mobile genetic elements (MGEs) such as IncA/C plasmids which can be transferred among bacteria. It is thought that AR Salmonella originates in food animals and is transmitted through food to humans. However, some AR Salmonella isolated from humans in the U.S. have different AR elements than those isolated from food animals, suggesting a different etiology for some AR human infections. The AR mechanisms identified in isolates from outside the U.S. are also predominantly different. For example the extended spectrum ?-lactamases (ESBLs) are found in human and animal isolates globally; however, in the U.S., ESBLs thus far have only been found in human and not food animal isolates. Commensal bacteria in animals including Escherichia coli and Enterococcus spp. may be reservoirs for AR mechanisms. Many of the AR genes and MGEs found in E. coli isolated from U.S. animals are similar to those found in Salmonella. Enterococcus spp. isolated from animals frequently carry MGEs with AR genes, including resistances to aminoglycosides (e.g., alleles of aac, ant, and aph), macrolides [e.g., erm(A), erm(B), and msrC], and tetracyclines [e.g., tet(K), (L), (M), (O), (S)]. Continuing investigations are required to help understand and mitigate the impact of AR bacteria on human and animal health. PMID:23734150
Blue Tigers, Black Tapirs, & the Pied Raven of the Faroe Islands: Teaching Genetic Drift Using Real-Life Animal Examples
ERIC Educational Resources Information Center
Robischon, Marcel
Genetic drift is a concept of population genetics that is central to understanding evolutionary processes and aspects of conservation biology. It is frequently taught using rather abstract representations. I introduce three real-life zoological examples, based on historical and recent color morphs of tigers, tapirs, and ravens, that can complementģ
Biochemical and genetic engineering of diatoms for polyunsaturated fatty acid biosynthesis.
Li, Hong-Ye; Lu, Yang; Zheng, Jian-Wei; Yang, Wei-Dong; Liu, Jie-Sheng
The role of diatoms as a source of bioactive compounds has been recently explored. Diatom cells store a high amount of fatty acids, especially certain polyunsaturated fatty acids (PUFAs). However, many aspects of diatom metabolism and the production of PUFAs remain unclear. This review describes a number of technical strategies, such as modulation of environmental factors (temperature, light, chemical composition of culture medium) and culture methods, to influence the content of PUFAs in diatoms. Genetic engineering, a newly emerging field, also plays an important role in controlling the synthesis of fatty acids in marine microalgae. Several key points in the biosynthetic pathway of PUFAs in diatoms as well as recent progresses are also a critical part and are summarized here. PMID:24402175
Biochemical and Genetic Engineering of Diatoms for Polyunsaturated Fatty Acid Biosynthesis
Li, Hong-Ye; Lu, Yang; Zheng, Jian-Wei; Yang, Wei-Dong; Liu, Jie-Sheng
The role of diatoms as a source of bioactive compounds has been recently explored. Diatom cells store a high amount of fatty acids, especially certain polyunsaturated fatty acids (PUFAs). However, many aspects of diatom metabolism and the production of PUFAs remain unclear. This review describes a number of technical strategies, such as modulation of environmental factors (temperature, light, chemical composition of culture medium) and culture methods, to influence the content of PUFAs in diatoms. Genetic engineering, a newly emerging field, also plays an important role in controlling the synthesis of fatty acids in marine microalgae. Several key points in the biosynthetic pathway of PUFAs in diatoms as well as recent progresses are also a critical part and are summarized here. PMID:24402175
Biochemical and genetic engineering strategies to enhance hydrogen production in photosynthetic algae and cyanobacteria.
Srirangan, Kajan; Pyne, Michael E; Perry Chou, C
As an energy carrier, hydrogen gas is a promising substitute to carbonaceous fuels owing to its superb conversion efficiency, non-polluting nature, and high energy content. At present, hydrogen is predominately synthesized via chemical reformation of fossil fuels. While various biological methods have been extensively explored, none of them is justified as economically feasible. A sustainable platform for biological production of hydrogen will certainly impact the biofuel market. Among a selection of biological systems, algae and cyanobacteria have garnered major interests as potential cell factories for hydrogen production. In conjunction with photosynthesis, these organisms utilize inexpensive inorganic substrates and solar energy for simultaneous biosynthesis and hydrogen evolution. However, the hydrogen yield associated with these organisms remains far too low to compete with the existing chemical systems. This article reviews recent advances of biochemical, bioprocess, and genetic engineering strategies in circumventing technological limitations to hopefully improve the applicative potential of these photosynthetic hydrogen production systems. PMID:21514821
Genetic engineering of Trichoderma to produce strains with novel cellulase profiles.
Harkki, A; Mõntylõ, A; Penttilõ, M; Muttilainen, S; B³hler, R; Suominen, P; Knowles, J; Nevalainen, H
Genetic engineering has been used to modify the proportion of different cellulases produced by a hypercellulolytic Trichoderma reesei mutant strain. A general expression vector, pAMH110, containing the promoter and terminator sequences of the strongly expressed main cellobiohydrolase 1 (cbh1) gene was used to overexpress a cDNA coding for EGI, the major endoglucanase (1,4,beta-D-glucan glucanohydrolase, EC 18.104.22.168). An in vitro modified cbh1 cDNA, incapable of coding for active enzyme, was used to inactivate the major cellobiohydrolase (1,4-beta-D-glucan cellobiohydrolase, EC 220.127.116.11) gene. In this way, new strains producing elevated amounts of the specific endoglucanase 1 (EGI) and/or lacking the major cellobiohydrolase (CBHI) were produced, and these have been further characterized. PMID:1367030
Production of 3-hydroxy-n-phenylalkanoic acids by a genetically engineered strain of Pseudomonas putida.
Sandoval, Angel; Arias-Barrau, Elsa; Bermejo, Francisco; Ca±edo, Librada; Naharro, Germßn; Olivera, ElĒas R; Luengo, Josķ M
Overexpression of the gene encoding the poly-3-hydroxy-n-phenylalkanoate (PHPhA) depolymerase (phaZ) in Pseudomonas putida U avoids the accumulation of these polymers as storage granules. In this recombinant strain, the 3-OH-acyl-CoA derivatives released from the different aliphatic or aromatic poly-3-hydroxyalkanoates (PHAs) are catabolized through the beta-oxidation pathway and transformed into general metabolites (acetyl-CoA, succinyl-CoA, phenylacetyl-CoA) or into non-metabolizable end-products (cinnamoyl-CoA). Taking into account the biochemical, pharmaceutical and industrial interest of some PHA catabolites (i.e., 3-OH-PhAs), we designed a genetically engineered strain of P. putida U (P. putida U DeltafadBA-phaZ) that efficiently bioconverts (more than 80%) different n-phenylalkanoic acids into their 3-hydroxyderivatives and excretes these compounds into the culture broth. PMID:15800732
Heritable multiplex genetic engineering in rats using CRISPR/Cas9.
Ma, Yuanwu; Shen, Bin; Zhang, Xu; Lu, Yingdong; Chen, Wei; Ma, Jing; Huang, Xingxu; Zhang, Lianfeng
The CRISPR/Cas9 system has been proven to be an efficient gene-editing tool for genome modification of cells and organisms. Multiplex genetic engineering in rat holds a bright future for the study of complex disease. Here, we show that this system enables the simultaneous disruption of four genes (ApoE, B2m, Prf1, and Prkdc) in rats in one-step, by co-injection of Cas9 mRNA and sgRNAs into fertilized eggs. We further observed the gene modifications are germline transmittable, and confirmed the off-target mutagenesis and mosaicism are rarely detected by comprehensive analysis. Thus, the CRISPR/Cas9 system makes it possible to efficiently and reliably generate gene knock-out rats. PMID:24598943
Decontamination of prion protein (BSE301V) using a genetically engineered protease.
Dickinson, J; Murdoch, H; Dennis, M J; Hall, G A; Bott, R; Crabb, W D; Penet, C; Sutton, J M; Raven, N D H
A previous study has demonstrated the potential of alkaline proteases to inactivate bovine spongiform encephalopathy (BSE301V). Here we explored the use of MC3, a genetically engineered variant of Bacillus lentus subtilisin. MC3 was used to digest BSE301V infectious mouse brain homogenate (iMBH). MC3 eliminated all detectable 6H4-immunoreactive material at pH 10 and 12; however, Proteinase K was only partially effective at pH 12. When bioassayed in VM mice, MC3- and Proteinase K-digested iMBH gave respectively 66.6% and 22.7% survival rates. Using a titration series for disease incubation, this equates to a >7log reduction in infectivity for MC3 and >6log reduction for Proteinase K. This study demonstrates the potential for thermostable proteases to be developed as effective inactivation processes for prion agents in healthcare management. PMID:19201054
Different methods of selecting animals for genotyping to maximize the amount of genetic information known in the population
Technology Transfer Automated Retrieval System (TEKTRAN)
A simulation study was carried out to develop and compare different methods of sampling animals to be genotyped. The simulated pedigrees included 5,000 animals and were assigned genotypes based on assumed allelic frequencies (favorable/unfavorable) of 0.3/0.7, 0.5/0.5, and 0.8/0.2. A real beef cat...
Biochemical, genetic, and metabolic engineering strategies to enhance coproduction of 1-propanol and ethanol in engineered Escherichia coli.
Srirangan, Kajan; Liu, Xuejia; Westbrook, Adam; Akawi, Lamees; Pyne, Michael E; Moo-Young, Murray; Chou, C Perry
We recently reported the heterologous production of 1-propanol in Escherichia coli via extended dissimilation of succinate under anaerobic conditions through expression of the endogenous sleeping beauty mutase (Sbm) operon. In the present work, we demonstrate high-level coproduction of 1-propanol and ethanol by developing novel engineered E. coli strains with effective cultivation strategies. Various biochemical, genetic, metabolic, and physiological factors affecting relative levels of acidogenesis and solventogenesis during anaerobic fermentation were investigated. In particular, CPC-PrOH3, a plasmid-free propanogenic E. coli strain derived by activating the Sbm operon on the genome, showed high levels of solventogenesis accounting for up to 85 % of dissimilated carbon. Anaerobic fed-batch cultivation of CPC-PrOH3 with glycerol as the major carbon source produced high titers of nearly 7 g/L 1-propanol and 31 g/L ethanol, implying its potential industrial applicability. The activated Sbm pathway served as an ancillary channel for consuming reducing equivalents upon anaerobic dissimilation of glycerol, resulting in an enhanced glycerol dissimilation and a major metabolic shift from acidogenesis to solventogenesis. PMID:25301579
Dr. Campbell's Bio111 Exam #4 Spring 2007 Biology 111 In-Class Exam #4 Cancer, HIV, & Genetic Engineering
Campbell, A. Malcolm
Dr. Campbell's Bio111 Exam #4 Ł Spring 2007 1 Biology 111 In-Class Exam #4 Ł Cancer, HIV, & Genetic Engineering There is no time limit on this test, though I have tried to design one that you should be able;Dr. Campbell's Bio111 Exam #4 Ł Spring 2007 3 6 pts. 5) List the main receptors on human cells used
Development of genetically engineered human intestinal cells for regulated insulin secretion using rAAV-mediated gene transfer
Shiue-Cheng Tang; Athanassios Sambanis
Cell-based therapies for treating insulin-dependent diabetes (IDD) can provide a more physiologic regulation of blood glucose levels in a less invasive fashion than daily insulin injections. Promising cells include intestinal enteroendocrine cells genetically engineered to secrete insulin in response to physiologic stimuli; responsiveness occurs at the exocytosis level to regulate the acute release of recombinant insulin. In this work, we
Surrogate species selection for assessing potential adverse environmental impacts of genetically engineered plants on non-target organisms
Technology Transfer Automated Retrieval System (TEKTRAN)
Most regulatory authorities require that developers of genetically engineered insect-resistant (GEIR) crops evaluate the potential for these crops to have adverse impacts on valued non-target organisms (NTOs), i.e., organisms not intended to be controlled by the trait. In many cases, impacts to NTOs...
Expression of Engineered Nuclear Male Sterility in Brassica napus (Genetics, Morphology, Cytology, and Sensitivity to Temperature).
Denis, M.; Delourme, R.; Gourret, J. P.; Mariani, C.; Renard, M.
A dominant genetic male sterility trait obtained through transformation in rapeseed (Brassica napus) was studied in the progenies of 11 transformed plants. The gene conferring the male sterility consists of a ribonuclease gene under the control of a tapetum-specific promoter. Two ribonuclease genes, RNase T1 and barnase, were used. The chimaeric ribonuclease gene was linked to the bialophos-resistance gene, which confers resistance to the herbicide phosphinotricine (PPT). The resistance to the herbicide was used as a dominant marker for the male sterility trait. The study presented here concerns three aspects of this engineered male sterility: genetics correlated with the segregation of the T-DNA in the progenies; expression of the male sterility in relation to the morphology and cytology of the androecium; and stability of the engineered male sterility under different culture conditions. Correct segregation, 50% male-sterile, PPT-resistant plants, and 50% male-fertile, susceptible plants were observed in the progeny of seven transformants. The most prominent morphological change in the male-sterile flowers was a noticeable reduction in the length of the stamen filament. The first disturbances of microsporogenesis were observed from the free microspore stage and were followed by a simultaneous degeneration of microspore and tapetal cell content. At anthesis, the sterile anthers contained only empty exines. In some cases, reversion to fertility of male-sterile plants has been observed. Both ribonuclease genes are susceptible to instability. Instability of the RNase T1-male sterility trait increased at temperatures higher than 25[deg] C. Our results do not allow us to confirm this observation for the barnase male-sterile plants. However, the male-sterile plants of the progeny of two independent RNase T1 transformants were stably male sterile under all conditions studied. PMID:12231785
Examining strategies to facilitate vitamin B1 biofortification of plants by genetic engineering.
Pourcel, Lucille; Moulin, Michael; Fitzpatrick, Teresa B
Thiamin (vitamin B1) is made by plants and microorganisms but is an essential micronutrient in the human diet. All organisms require it as a cofactor in its form as thiamin pyrophosphate (TPP) for the activity of key enzymes of central metabolism. In humans, deficiency is widespread particularly in populations where polished rice is a major component of the diet. Considerable progress has been made on the elucidation of the biosynthesis pathway within the last few years enabling concrete strategies for biofortification purposes to be devised, with a particular focus here on genetic engineering. Furthermore, the vitamin has been shown to play a role in both abiotic and biotic stress responses. The precursors for de novo biosynthesis of thiamin differ between microorganisms and plants. Bacteria use intermediates derived from purine and isoprenoid biosynthesis, whereas the pathway in yeast involves the use of compounds from the vitamin B3 and B6 groups. Plants on the other hand use a combination of the bacterial and yeast pathways and there is subcellular partitioning of the biosynthesis steps. Specifically, thiamin biosynthesis occurs in the chloroplast of plants through the separate formation of the pyrimidine and thiazole moieties, which are then coupled to form thiamin monophosphate (TMP). Phosphorylation of thiamin to form TPP occurs in the cytosol. Therefore, thiamin (or TMP) must be exported from the chloroplast to the cytosol for the latter step to be executed. The regulation of biosynthesis is mediated through riboswitches, where binding of the product TPP to the pre-mRNA of a biosynthetic gene modulates expression. Here we examine and hypothesize on genetic engineering approaches attempting to increase the thiamin content employing knowledge gained with the model plant Arabidopsis thaliana. We will discuss the regulatory steps that need to be taken into consideration and can be used a prerequisite for devising such strategies in crop plants. PMID:23755056
Development of New Modular Genetic Tools for Engineering the Halophilic Archaeon Halobacterium salinarum
Silva-Rocha, Rafael; Pontelli, Marjorie Cornejo; Furtado, Gilvan Pessoa; Zaramela, Livia Soares; Koide, Tie
Our ability to genetically manipulate living organisms is usually constrained by the efficiency of the genetic tools available for the system of interest. In this report, we present the design, construction and characterization of a set of four new modular vectors, the pHsal series, for engineering Halobacterium salinarum, a model halophilic archaeon widely used in systems biology studies. The pHsal shuttle vectors are organized in four modules: (i) the E. coliÆs specific part, containing a ColE1 origin of replication and an ampicillin resistance marker, (ii) the resistance marker and (iii) the replication origin, which are specific to H. salinarum and (iv) the cargo, which will carry a sequence of interest cloned in a multiple cloning site, flanked by universal M13 primers. Each module was constructed using only minimal functional elements that were sequence edited to eliminate redundant restriction sites useful for cloning. This optimization process allowed the construction of vectors with reduced sizes compared to currently available platforms and expanded multiple cloning sites. Additionally, the strong constitutive promoter of the fer2 gene was sequence optimized and incorporated into the platform to allow high-level expression of heterologous genes in H. salinarum. The system also includes a new minimal suicide vector for the generation of knockouts and/or the incorporation of chromosomal tags, as well as a vector for promoter probing using a GFP gene as reporter. This new set of optimized vectors should strongly facilitate the engineering of H. salinarum and similar strategies could be implemented for other archaea. PMID:26061363
Governing the moral economy: Animal engineering, ethics and the liberal government of science
Harvey, Alison; Salter, Brian
The preferred Western model for science governance has come to involve attending to the perspectives of the public. In practice, however, this model has been criticised for failing to promote democracy along participatory lines. We argue that contemporary approaches to science policy making demonstrate less the failure of democracy and more the success of liberal modes of government in adapting to meet new governance challenges. Using a case study of recent UK policy debates on scientific work mixing human and animal biological material, we show first how a æmoral economyÆ is brought into being as a regulatory domain and second how this domain is governed to align cultural with scientific values. We suggest that it is through these practices that the state assures its aspirations for enhancing individual and collective prosperity through technological advance are met. PMID:22507952
Governing the moral economy: animal engineering, ethics and the liberal government of science.
Harvey, Alison; Salter, Brian
The preferred Western model for science governance has come to involve attending to the perspectives of the public. In practice, however, this model has been criticised for failing to promote democracy along participatory lines. We argue that contemporary approaches to science policy making demonstrate less the failure of democracy and more the success of liberal modes of government in adapting to meet new governance challenges. Using a case study of recent UK policy debates on scientific work mixing human and animal biological material, we show first how a 'moral economy' is brought into being as a regulatory domain and second how this domain is governed to align cultural with scientific values. We suggest that it is through these practices that the state assures its aspirations for enhancing individual and collective prosperity through technological advance are met. PMID:22507952
Detection of Anthrax Toxin in the Serum of Animals Infected with Bacillus anthracis by Using Engineered Immunoassays
Mabry, Robert; Brasky, Kathleen; Geiger, Robert; Carrion, Ricardo; Hubbard, Gene B.; Leppla, Stephen; Patterson, Jean L.; Georgiou, George; Iverson, B. L.
Several strategies that target anthrax toxin are being developed as therapies for infection by Bacillus anthracis. Although the action of the tripartite anthrax toxin has been extensively studied in vitro, relatively little is known about the presence of toxins during an infection in vivo. We developed a series of sensitive sandwich enzyme-linked immunosorbent assays (ELISAs) for detection of both the protective antigen (PA) and lethal factor (LF) components of the anthrax exotoxin in serum. The assays utilize as capture agents an engineered high-affinity antibody to PA, a soluble form of the extracellular domain of the anthrax toxin receptor (ANTXR2/CMG2), or PA itself. Sandwich immunoassays were used to detect and quantify PA and LF in animals infected with the Ames or Vollum strains of anthrax spores. PA and LF were detected before and after signs of toxemia were observed, with increasing levels reported in the late stages of the infection. These results represent the detection of free PA and LF by ELISA in the systemic circulation of two animal models exposed to either of the two fully virulent strains of anthrax. Simple anthrax toxin detection ELISAs could prove useful in the evaluation of potential therapies and possibly as a clinical diagnostic to complement other strategies for the rapid identification of B. anthracis infection. PMID:16760326
de Lange, W.J.; Hegge, L.F.; Grimes, A.C.; Tong, C.W.; Brost, T.M.; Moss, R.L.; Ralphe, J.C.
Rationale Cardiomyocytes cultured in a mechanically active three-dimensional configuration can be used for studies that correlate contractile performance to cellular physiology. Current engineered cardiac tissue (ECT) models employ cells derived from either rat or chick hearts. Development of a murine ECT would provide access to many existing models of cardiac disease, and open the possibility of performing targeted genetic manipulation with the ability to directly assess contractile and molecular variables. Objective To generate, characterize and validate mouse ECT using a physiologically relevant model of hypertrophic cardiomyopathy. Methods and Results We generated mechanically integrated ECT using isolated neonatal mouse cardiac cells derived from both wild type and myosin binding protein C (cMyBP-C) null mouse hearts. The murine ECT's produce consistent contractile forces that follow the Frank-Starling law, accept physiologic pacing. cMyBP-C null ECT show characteristic acceleration of contraction kinetics. Adenoviral-mediated expression of human cMyBP-C in murine cMyBP-C null ECT restores contractile properties to levels indistinguishable from those of wild type ECT. Importantly, the cardiomyocytes used to construct the cMyBP-C?/? ECT have yet to undergo the significant hypertrophic remodeling that occurs in vivo. Thus, this murine ECT model reveals a contractile phenotype that is specific to the genetic mutation rather than to secondary remodeling events. Conclusions Data presented here show mouse ECT to be an efficient and cost effective platform to study the primary effects of genetic manipulation on cardiac contractile function. This model provides a previously unavailable tool to study specific sarcomeric protein mutations in an intact mammalian muscle system. PMID:21566213
Frontiers of torenia research: innovative ornamental traits and study of ecological interaction networks through genetic engineering
Advances in research in the past few years on the ornamental plant torenia (Torenia spps.) have made it notable as a model plant on the frontier of genetic engineering aimed at studying ornamental characteristics and pest control in horticultural ecosystems. The remarkable advantage of torenia over other ornamental plant species is the availability of an easy and high-efficiency transformation system for it. Unfortunately, most of the current torenia research is still not very widespread, because this species has not become prominent as an alternative to other successful model plants such as Arabidopsis, snapdragon and petunia. However, nowadays, a more global view using not only a few selected models but also several additional species are required for creating innovative ornamental traits and studying horticultural ecosystems. We therefore introduce and discuss recent research on torenia, the family Scrophulariaceae, for secondary metabolite bioengineering, in which global insights into horticulture, agriculture and ecology have been advanced. Floral traits, in torenia particularly floral color, have been extensively studied by manipulating the flavonoid biosynthetic pathways in flower organs. Plant aroma, including volatile terpenoids, has also been genetically modulated in order to understand the complicated nature of multi-trophic interactions that affect the behavior of predators and pollinators in the ecosystem. Torenia would accordingly be of great use for investigating both the variation in ornamental plants and the infochemical-mediated interactions with arthropods. PMID:23803155
Pathway engineering for phenolic acid accumulations in Salvia miltiorrhiza by combinational genetic manipulation.
Zhang, Yuan; Yan, Ya-Ping; Wu, Yu-Cui; Hua, Wen-Ping; Chen, Chen; Ge, Qian; Wang, Zhe-Zhi
To produce beneficial phenolic acids for medical and commercial purposes, researchers are interested in improving the normally low levels of salvianolic acid B (Sal B) produced by Salvia miltiorrhiza. Here, we present a strategy of combinational genetic manipulation to enrich the precursors available for Sal B biosynthesis. This approach, involving the lignin pathway, requires simultaneous, ectopic expression of an Arabidopsis Production of Anthocyanin Pigment 1 transcription factor (AtPAP1) plus co-suppression of two endogenous, key enzyme genes: cinnamoyl-CoA reductase (SmCCR) and caffeic acid O-methyltransferase (SmCOMT). Compared with the untransformed control, we achieved a greater accumulation of Sal B (up to 3-fold higher) along with a reduced lignin concentration. This high-Sal B phenotype was stable in roots during vegetative growth and was closely correlated with increased antioxidant capacity for the corresponding plant extracts. Although no outward change in phenotype was apparent, we characterized the molecular phenotype through integrated analysis of transcriptome and metabolome profiling. Our results demonstrated the far-reaching consequences of phenolic pathway perturbations on carbohydrate metabolism, respiration, photo-respiration, and stress responses. This report is the first to describe the production of valuable end products through combinational genetic manipulation in S. miltiorrhiza plants. Our strategy will be effective in efforts to metabolically engineer multi-branch pathway(s), such as the phenylpropanoid pathway, in economically significant medicinal plants. PMID:24269612
Camelina as a sustainable oilseed crop: contributions of plant breeding and genetic engineering.
Vollmann, Johann; Eynck, Christina
Camelina is an underutilized Brassicaceae oilseed plant with a considerable agronomic potential for biofuel and vegetable oil production in temperate regions. In contrast to most Brassicaceae, camelina is resistant to alternaria black spot and other diseases and pests. Sequencing of the camelina genome revealed an undifferentiated allohexaploid genome with a comparatively large number of genes and low percentage of repetitive DNA. As there is a close relationship between camelina and the genetic model plant Arabidopsis, this review aims at exploring the potential of translating basic Arabidopsis results into a camelina oilseed crop for food and non-food applications. Recently, Arabidopsis genes for drought resistance or increased photosynthesis and overall productivity have successfully been expressed in camelina. In addition, gene constructs affecting lipid metabolism pathways have been engineered into camelina for synthesizing either long-chain polyunsaturated fatty acids, hydroxy fatty acids or high-oleic oils in particular camelina strains, which is of great interest in human food, industrial or biofuel applications, respectively. These results confirm the potential of camelina to serve as a biotechnology platform in biorefinery applications thus justifying further investment in breeding and genetic research for combining agronomic potential, unique oil quality features and biosafety into an agricultural production system. PMID:25706640
NSDL National Science Digital Library
Jennifer Doherty
This activity helps students to understand basic principles of genetics, including relationships of genotype to phenotype, concepts of recessive and dominant alleles, and how understanding meiosis and fertilization provides the basis for understanding inheritance, as summarized in Punnett squares. The Student Handout includes an analysis of the inheritance of albinism that teaches all of these concepts, a Coin Toss Genetics activity that helps students understand the probabilistic nature of Punnett square predictions, and an analysis of the inheritance of sickle cell anemia that reinforces the basic concepts and introduces some of the complexities of genetics. The Genetics Supplement includes two additional activities, an analysis of student data on the sex makeup of sibships and pedigree analyses of recessive and dominant alleles with challenge questions that introduce the role of mutations and an evaluation of Punnett squares and pedigrees as models of inheritance.
Research review paper The impact of genetic modification of human foods in the 21st century: A review
Stella G. Uzogara
Genetic engineering of food is the science which involves deliberate modification of the genetic material of plants or animals. It is an old agricultural practice carried on by farmers since early histor- ical times, but recently it has been improved by technology. Many foods consumed today are either genetically modified (GM) whole foods, or contain ingredients derived from gene modification
A prototype stable RNA identification cassette for monitoring plasmids of genetically engineered microorganisms
NASA Technical Reports Server (NTRS)
Hedenstierna, K. O.; Lee, Y. H.; Yang, Y.; Fox, G. E.
A prototype stable RNA identification cassette for monitoring genetically engineered plasmids carried by strains of Escherichia coli has been developed. The cassette consists of a Vibrio proteolyticus 5S ribosomal RNA (rRNA) gene surrounded by promoters and terminators from the rrnB operon of Escherischia coli. The identifier RNA is expressed and successfully processed so that approximately 30% of the 5S rRNA isolated from either whole cells or 70S ribosomes is of the V. proteolyticus type. Cells carrying the identifier are readily detectable by hybridization. Accurate measurements show that the identification cassette has little effect on fitness compared to a strain containing an analogous plasmid carrying wild type E. coli 5S rRNA, and the V. proteolyticus 5S rRNA gene is not inactivated after prolonged growth. These results demonstrate the feasibility of developing small standardized identification cassettes that can utilize already existing highly sensitive rRNA detection methods. Cassettes of this type could in principle be incorporated into either the engineered regions of recombinant plasmids or their hosts.
Two concepts of dignity for humans and non-human organisms in the context of genetic engineering
Philipp Balzer; Klaus Peter Rippe; Peter Schaber
The 1992 incorporation of an article by referendum in the Swiss Constitution mandating that the federal government issue regulations\\u000a on the use of genetic material that take into account the dignity of nonhuman organism raises philosophical questions about\\u000a how we should understand what is meant by ōthe dignity of nonhuman animals,ö and about what sort of moral demands arise from
A Compendium of the Mouse Mammary Tumor Biologist: From the Initial Observations in the House Mouse to the Development of Genetically Engineered Mice
Cardiff, Robert D.; Kenney, Nicholas
For over a century, mouse mammary tumor biology and the associated mouse mammary tumor virus (MMTV) have served as the foundation for experimental cancer research, in general, and, in particular, experimental breast cancer research. Spontaneous mouse mammary tumors were the basis for studies of the natural history of neoplasia, oncogenic viruses, host responses, endocrinology and neoplastic progression. However, lacking formal proof of a human mammary tumor virus, the preeminence of the mouse model faded in the 1980s. Since the late 1980s, genetically engineered mice (GEM) have proven extremely useful for studying breast cancer and have become the animal model for human breast cancer. Hundreds of mouse models of human breast cancer have been developed since the first demonstration in 1984. The GEM have attracted a new generation of molecular and cellular biologists eager to apply their skill sets to these surrogates of the human disease. Newcomers often enter the field without an appreciation of the origins of mouse mammary tumor biology and the basis for many of the prevailing concepts. Our purpose in writing this compendium is to extend an ōolive branchö while simultaneously deepen the knowledge of the novice mouse mammary tumor biologist as they journey into a field rich in pathology and genetics spanning several centuries. PMID:20961975
Ex situ conservation genetics: a review of molecular studies on the genetic consequences of captive breeding programmes for endangered animal species
Kathrin A. Witzenberger; Axel Hochkirch
Captive breeding has become an important tool in species conservation programmes. Current management strategies for ex situ populations are based on theoretical models, which have mainly been tested in model species or assessed using studbook data.\\u000a During recent years an increasing number of molecular genetic studies have been published on captive populations of several\\u000a endangered species. However, a comprehensive analysis
Potential Large Animal Models for Gene Therapy of Human Genetic Diseases of Immune and Blood Cell Systems
Thomas R. Bauer; Rima L. Adler; Dennis D. Hickstein
Genetic mutations involving the cellular components of the hematopoietic systemŚred blood cells, white blood cells, and plateletsŚmanifest clinically as anemia, infection, and bleed- ing. Although gene targeting has recapitulated many of these diseases in mice, these murine homologues are limited as translational models by their small size and brief life span as well as the fact that mutations induced by
U.S. Unit Opens Way to Patent Animals; Humans Seen Likely to Be Next Test Case.
ERIC Educational Resources Information Center
Wheeler, David L.
With a decision on an oyster developed at the University of Washington, the federal Board of Patent Appeals and Interferences has opened the way to granting patents for animals and animal improvements developed through genetic engineering and other scientific methods. (MSE)
The genetic heterogeneity and mutational burden of engineered melanomas in zebrafish models
Background Melanoma is the most deadly form of skin cancer. Expression of oncogenic BRAF or NRAS, which are frequently mutated in human melanomas, promote the formation of nevi but are not sufficient for tumorigenesis. Even with germline mutated p53, these engineered melanomas present with variable onset and pathology, implicating additional somatic mutations in a multi-hit tumorigenic process. Results To decipher the genetics of these melanomas, we sequence the protein coding exons of 53 primary melanomas generated from several BRAFV600E or NRASQ61K driven transgenic zebrafish lines. We find that engineered zebrafish melanomas show an overall low mutation burden, which has a strong, inverse association with the number of initiating germline drivers. Although tumors reveal distinct mutation spectrums, they show mostly C?>?T transitions without UV light exposure, and enrichment of mutations in melanogenesis, p53 and MAPK signaling. Importantly, a recurrent amplification occurring with pre-configured drivers BRAFV600E and p53-/- suggests a novel path of BRAF cooperativity through the protein kinase A pathway. Conclusion This is the first analysis of a melanoma mutational landscape in the absence of UV light, where tumors manifest with remarkably low mutation burden and high heterogeneity. Genotype specific amplification of protein kinase A in cooperation with BRAF and p53 mutation suggests the involvement of melanogenesis in these tumors. This work is important for defining the spectrum of events in BRAF or NRAS driven melanoma in the absence of UV light, and for informed exploitation of models such as transgenic zebrafish to better understand mechanisms leading to human melanoma formation. PMID:24148783
Animal Genotype x Environment Interactions
Technology Transfer Automated Retrieval System (TEKTRAN)
While genetics define the potential of an animal, the expression of those genetics is often modified by the environment in which the animal exists. Environmental effects may modify genetic expression of a trait uniformly across all possible genotypes or the amount of modification may be specific to ...
Different methods of selecting animals for genotyping to maximize the amount of genetic information known in the population.
Technology Transfer Automated Retrieval System (TEKTRAN)
It is possible to predict genotypes of some individuals based on genotypes of relatives. Different methods of sampling individuals to be genotyped from populations were evaluated using simulation. Simulated pedigrees included 5,000 animals and were assigned genotypes based on assumed allelic frequ...
NSDL National Science Digital Library
This animation demonstrates the stages of mitosis in an animal cell. Use the control buttons in the upper left to run the complete animation. Click on any intermediate stage (for example, Anaphase), and see a representative still frame.
Biology 2250 Principles of Genetics
Innes, David J.
? - Genetics is the study of heredity and variation - Examples of genetic variation 1. Domesticated species 2 American J. Human Genetics Heredity Annals of Human Genetics Hereditas Opthalmic Genetics Japanese Journal of Human Genetics Human Genetics Journal of Heredity CurrentGenetics Molecular Biology and Evolution Animal
Biological detoxification of the mycotoxin deoxynivalenol and its use in genetically engineered crops and feed additives.
Karlovsky, Petr
Deoxynivalenol (DON) is the major mycotoxin produced by Fusarium fungi in grains. Food and feed contaminated with DON pose a health risk to humans and livestock. The risk can be reduced by enzymatic detoxification. Complete mineralization of DON by microbial cultures has rarely been observed and the activities turned out to be unstable. The detoxification of DON by reactions targeting its epoxide group or hydroxyl on carbon 3 is more feasible. Microbial strains that de-epoxidize DON under anaerobic conditions have been isolated from animal digestive system. Feed additives claimed to de-epoxidize trichothecenes enzymatically are on the market but their efficacy has been disputed. A new detoxification pathway leading to 3-oxo-DON and 3-epi-DON was discovered in taxonomically unrelated soil bacteria from three continents; the enzymes involved remain to be identified. Arabidopsis, tobacco, wheat, barley, and rice were engineered to acetylate DON on carbon 3. In wheat expressing DON acetylation activity, the increase in resistance against Fusarium head blight was only moderate. The Tri101 gene from Fusarium sporotrichioides was used; Fusarium graminearum enzyme which possesses higher activity towards DON would presumably be a better choice. Glycosylation of trichothecenes occurs in plants, contributing to the resistance of wheat to F. graminearum infection. Marker-assisted selection based on the trichothecene-3-O-glucosyltransferase gene can be used in breeding for resistance. Fungal acetyltransferases and plant glucosyltransferases targeting carbon 3 of trichothecenes remain promising candidates for engineering resistance against Fusarium head blight. Bacterial enzymes catalyzing oxidation, epimerization, and less likely de-epoxidation of DON may extend this list in future. PMID:21691789
Current status of regulating biotechnology-derived animals in Canada: animal health and food safety considerations.
Kochhar, H P S; Evans, B R
Development of an effective regulatory system for genetically engineered animals and their products has been the subject of increasing discussion among researchers, industry and policy developers, as well as the public. Since transgenesis and cloning are relatively new scientific techniques, transgenic animals are 'novel' organisms for which there is limited information. The issues associated with the regulation of transgenic animals pertain to environmental impact, human food safety, animal health and welfare, trade and ethics. It is a challenge for the developers to prove the safety of the products of biotechnology-derived animals and also for regulators to regulate this increasingly powerful technology with limited background information. In principle, an effective regulatory sieve should permit safe products while forming a formidable barrier for those posing an unacceptable risk. Regulatory initiatives for biotechnology-derived animals and their products should be able to ensure high standards for human and animal health, a sound scientific basis for evaluation; transparency and public involvement, and maintenance of genetic diversity. This review proposes a regulatory regime that is based on scientific risk based assessment and approval of products or by-products of biotechnology-derived animals and its application in context to Canadian regulations. PMID:17097725
Impact of a Genetically Engineered Bacterium with Enhanced Alkaline Phosphatase Activity on Marine Phytoplankton Communities
Sobecky, P. A.; Schell, M. A.; Moran, M. A.; Hodson, R. E.
An indigenous marine Achromobacter sp. was isolated from coastal Georgia seawater and modified in the laboratory by introduction of a plasmid with a phoA hybrid gene that directed constitutive overproduction of alkaline phosphatase. The effects of this "indigenous" genetically engineered microorganism (GEM) on phosphorus cycling were determined in seawater microcosms following the addition of a model dissolved organic phosphorus compound, glycerol 3-phosphate, at a concentration of 1 or 10 (mu)M. Within 48 h, a 2- to 10-fold increase in the concentration of inorganic phosphate occurred in microcosms containing the GEM (added at an initial density equivalent to 8% of the total bacterial population) relative to controls containing only natural microbial populations, natural populations with the unmodified Achromobacter sp., or natural populations with the Achromobacter sp. containing the plasmid but not the phoA gene. Secondary effects of the GEM on the phytoplankton community were observed after several days, evident as sustained increases in phytoplankton biomass (up to 14-fold) over that in controls. Even in the absence of added glycerol 3-phosphate, a numerically stable GEM population (averaging 3 to 5% of culturable bacteria) was established within 2 to 3 weeks of introduction into seawater. Moreover, alkaline phosphatase activity in microcosms with the GEM was substantially higher than that in controls for up to 25 days, and microcosms containing the GEM maintained the potential for net phosphate accumulation above control levels for longer than 1 month. PMID:16535222
Genetic engineering of Escherichia coli for the economical production of sialylated oligosaccharides.
Fierfort, Nicolas; Samain, Eric
We have previously described a microbiological process for the conversion of lactose into 3'sialyllactose and other ganglioside sugars by living Escherichia coli cells expressing the appropriate recombinant glycosyltransferase genes. In this system the activated sialic acid donor (CMP-Neu5Ac) was generated from exogenous sialic acid, which was transported into the cells by the permease NanT. Since sialic acid is an expensive compound, a more economical process has now been developed by genetically engineering E. coli K12 to be capable of generating CMP-Neu5Ac using its own internal metabolism. Mutant strains devoid of Neu5Ac aldolase and of ManNAc kinase were shown to efficiently produce 3'sialyllactose by coexpressing the alpha-2,3-sialyltransferase gene from Neisseria meningitidis with the neuC, neuB and neuACampylobacter jejuni genes encoding N-acetylglucosamine-6-phosphate-epimerase, sialic acid synthase and CMP-Neu5Ac synthetase, respectively. A sialyllactose concentration of 25 g l(-1) was obtained in long-term high cell density culture with a continuous lactose feed. This high concentration and low cost of fermentation medium should make possible to use sialylated oligosaccharides in new fields such as the food industry. PMID:18378033
Behavior of pollutant-degrading microorganisms in aquifers. Predictions for genetically engineered organisms
Krumme, M.L.; Egestorff, J.; Timmis, K.N.; Dwyer, D.F. (Natioanl Research Center for Biotechnology, Braunschweig (Germany)); Smith, R.L. (Geological Survey, Boulder, CO (United States)); Thiem, S.M.; Tiedje, J.M. (Michigan State Univ., East Lansing, MI (United States))
Bioremediation via environmental introductions of degradative microorganisms requires that the microbes survive in substantial numbers and effect an increase in the rate and extent of pollutant removal. Combined field and microcosm studies were used to assess these abilities for laboratory-grown bacteria. Following introduction into a contaminated aquifer, viable cells of Pseudomonas sp. B13 were present in the contaminant plume for 447 days; die-off was rapid in pristine areas. In aquifer microcosms, survival of B13 and FR120, a genetically engineered derivative of B13 having enhanced catabolic capabilities for substituted aromatics, was comparable to B13 field results; both bacteria degraded target pollutants in microcosms made with aquifer samples from the aerobic zone of the pollutant plume. Results suggest that field studies with nonrecombinant microorganisms may be coupled to laboratory studies with derivative strains to estimate their bioremediative efficacy. Furthermore, laboratory strains of bacteria can survive for extended periods of time in nature and thus may have important bioremediative applications. 33 refs., 4 figs., 1 tab.
Cyanobacterial defense mechanisms against foreign DNA transfer and their impact on genetic engineering.
Stucken, Karina; Koch, Robin; Dagan, Tal
Cyanobacteria display a large diversity of cellular forms ranging from unicellular to complex multicellular filaments or aggregates. Species in the group present a wide range of metabolic characteristics including the fixation of atmospheric nitrogen, resistance to extreme environments, production of hydrogen, secondary metabolites and exopolysaccharides. These characteristics led to the growing interest in cyanobacteria across the fields of ecology, evolution, cell biology and biotechnology. The number of available cyanobacterial genome sequences has increased considerably in recent years, with more than 140 fully sequenced genomes to date. Genetic engineering of cyanobacteria is widely applied to the model unicellular strains Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942. However the establishment of transformation protocols in many other cyanobacterial strains is challenging. One obstacle to the development of these novel model organisms is that many species have doubling times of 48 h or more, much longer than the bacterial models E. coli or B. subtilis. Furthermore, cyanobacterial defense mechanisms against foreign DNA pose a physical and biochemical barrier to DNA insertion in most strains. Here we review the various barriers to DNA uptake in the context of lateral gene transfer among microbes and the various mechanisms for DNA acquisition within the prokaryotic domain. Understanding the cyanobacterial defense mechanisms is expected to assist in the development and establishment of novel transformation protocols that are specifically suitable for this group. PMID:24510140
The first crop plant genetically engineered to release an insect pheromone for defence
Bruce, Toby J.A.; Aradottir, Gudbjorg I.; Smart, Lesley E.; Martin, Janet L.; Caulfield, John C.; Doherty, Angela; Sparks, Caroline A.; Woodcock, Christine M.; Birkett, Michael A.; Napier, Johnathan A.; Jones, Huw D.; Pickett, John A.
Insect pheromones offer potential for managing pests of crop plants. Volatility and instability are problems for deployment in agriculture but could be solved by expressing genes for the biosynthesis of pheromones in the crop plants. This has now been achieved by genetically engineering a hexaploid variety of wheat to release (E)-?-farnesene (E?f), the alarm pheromone for many pest aphids, using a synthetic gene based on a sequence from peppermint with a plastid targeting amino acid sequence, with or without a gene for biosynthesis of the precursor farnesyl diphosphate. Pure E?f was produced in stably transformed wheat lines with no other detectable phenotype but requiring targeting of the gene produced to the plastid. In laboratory behavioural assays, three species of cereal aphids were repelled and foraging was increased for a parasitic natural enemy. Although these studies show considerable potential for aphid control, field trials employing the single and double constructs showed no reduction in aphids or increase in parasitism. Insect numbers were low and climatic conditions erratic suggesting the need for further trials or a closer imitation, in the plant, of alarm pheromone release. PMID:26108150
Genetic engineering activates biosynthesis of aromatic fumaric acid amides in the human pathogen Aspergillus fumigatus.
Kalb, Daniel; Heinekamp, Thorsten; Lackner, Gerald; Scharf, Daniel H; Dahse, Hans-Martin; Brakhage, Axel A; Hoffmeister, Dirk
The Aspergillus fumigatus nonribosomal peptide synthetase FtpA is among the few of this species whose natural product has remained unknown. Both FtpA adenylation domains were characterized in vitro. Fumaric acid was identified as preferred substrate of the first and both l-tyrosine and l-phenylalanine as preferred substrates of the second adenylation domain. Genetically engineered A. fumigatus strains expressed either ftpA or the regulator gene ftpR, encoded in the same cluster of genes, under the control of the doxycycline-inducible tetracycline-induced transcriptional activation (tet-on) cassette. These strains produced fumaryl-l-tyrosine and fumaryl-l-phenylalanine which were identified by liquid chromatography and high-resolution mass spectrometry. Modeling of the first adenylation domain in silico provided insight into the structural requirements to bind fumaric acid as peptide synthetase substrate. This work adds aromatic fumaric acid amides to the secondary metabolome of the important human pathogen A. fumigatus which was previously not known as a producer of these compounds. PMID:25527545
Evaluating oversight systems for emerging technologies: a case study of genetically engineered organisms.
Kuzma, Jennifer; Najmaie, Pouya; Larson, Joel
The U.S. oversight system for genetically engineered organisms (GEOs) was evaluated to develop hypotheses and derive lessons for oversight of other emerging technologies, such as nanotechnology. Evaluation was based upon quantitative expert elicitation, semi-standardized interviews, and historical literature analysis. Through an interdisciplinary policy analysis approach, blending legal, ethical, risk analysis, and policy sciences viewpoints, criteria were used to identify strengths and weaknesses of GEOs oversight and explore correlations among its attributes and outcomes. From the three sources of data, hypotheses and broader conclusions for oversight were developed. Our analysis suggests several lessons for oversight of emerging technologies: the importance of reducing complexity and uncertainty in oversight for minimizing financial burdens on small product developers; consolidating multi-agency jurisdictions to avoid gaps and redundancies in safety reviews; consumer benefits for advancing acceptance of GEO products; rigorous and independent pre- and post-market assessment for environmental safety; early public input and transparency for ensuring public confidence; and the positive role of public input in system development, informed consent, capacity, compliance, incentives, and data requirements and stringency in promoting health and environmental safety outcomes, as well as the equitable distribution of health impacts. Our integrated approach is instructive for more comprehensive analyses of oversight systems, developing hypotheses for how features of oversight systems affect outcomes, and formulating policy options for oversight of future technological products, especially nanotechnology products. PMID:20122100
Solute diffusion in genetically engineered silk-elastinlike protein polymer hydrogels.
Dinerman, Adam A; Cappello, Joseph; Ghandehari, Hamidreza; Hoag, Stephen W
The partitioning and diffusion behavior of theophylline, vitamin B(12), and cytochrome c in physically crosslinked networks of a genetically engineered silk-elastinlike protein-based (SELP) copolymer with an amino acid sequence of [(GVGVP)(4)GKGVP(GVGVP)(3)(GAGAGS)(4)](12) was investigated. The effect of gelation kinetics on the equilibrium swelling ratio and normalized dimensions of loaded SELP hydrogel disks before and after release studies was also examined. Size dependent release behavior was quantified by diffusion studies with equilibrium loaded SELP hydrogels. Direct loading diffusion studies confirmed that hydrogels produced by direct incorporation of cytochrome c with the aqueous SELP solution did not significantly influence the release behavior compared to equilibrium loaded hydrogels. An overall increase in the equilibrium swelling ratio after the release studies was observed. Analysis of the hydrogel disk dimensions after the release studies revealed no expansion of the disk dimensions. The apparent increase in the equilibrium swelling ratio was most likely due to a decrease in the hydrogel crosslinking density following the removal of the polymer soluble fraction over the course of the release study. PMID:12175743
Swelling behavior of a genetically engineered silk-elastinlike protein polymer hydrogel.
Dinerman, Adam A; Cappello, Joseph; Ghandehari, Hamidreza; Hoag, Stephen W
The influence of environmental conditions such as pH, temperature, and ionic strength on the equilibrium swelling ratio of physically crosslinked networks of a genetically engineered silk-elastinlike protein-based copolymer (SELP) with an amino acid repeat sequence of [(GVGVP)4GKGVP(GVGVP)3(GAGAGS)4]12 was investigated. The effects of gelation cure time and initial polymer concentration on the equilibrium swelling ratio and soluble fraction of the hydrogels were also studied. It was found that the soluble fraction linearly correlated with the initial polymer concentration at higher gelation times. Soluble fraction results suggest that final hydrogel water content may be controlled by both initial polymer concentration and gelation time. Equilibrium swelling studies demonstrated that these hydrogels are relatively insensitive to environmental changes such as pH, temperature, and ionic strength. Over the concentration range studied, it was found that an increase in gelation time at 37 degrees C resulted in lower hydrogel weight equilibrium swelling ratios, which corresponds to less soluble polymer released post-gelation. Together, these results have implications for the controlled delivery of bioactive agents from silk-elastinlike hydrogels. PMID:12194523
The private sector's role in public sector genetically engineered crop projects.
Potrykus, Ingo
There is widespread interest within academia to work on public good genetically engineered (GE) projects to the benefit of the poor, especially to use GE-technology to contribute to food security. Not a single product from this work has reached the market. The major cause is GE-regulation, which prevents use of the technology for public good beyond proof-of-concept (Potrykus, I. (2010) Lessons from the Humanitarian Golden Rice project: Regulation prevents development of public good GE-products (these Proceedings)). There is, however, another key problem responsible for the lack of deployment of public good GE-plants: the public sector is incompetent and disinterested for work beyond proof-of-concept, and has neither capability nor funding to develop GE-plant products and introduce them to growers and consumers. The private sector has the expertise for both and in the right circumstances can be ready to support the public sector in public good enterprises. Public-private-partnerships are the best solution so far, to advance exploitation of GE-technology to the benefit of the poor. Public-private-partnerships are viable, however, only, if there is mutual interest from the private sector and initiative and funding from the public sector. PMID:20637908
The first crop plant genetically engineered to release an insect pheromone for defence.
Bruce, Toby J A; Aradottir, Gudbjorg I; Smart, Lesley E; Martin, Janet L; Caulfield, John C; Doherty, Angela; Sparks, Caroline A; Woodcock, Christine M; Birkett, Michael A; Napier, Johnathan A; Jones, Huw D; Pickett, John A
Insect pheromones offer potential for managing pests of crop plants. Volatility and instability are problems for deployment in agriculture but could be solved by expressing genes for the biosynthesis of pheromones in the crop plants. This has now been achieved by genetically engineering a hexaploid variety of wheat to release (E)-?-farnesene (E?f), the alarm pheromone for many pest aphids, using a synthetic gene based on a sequence from peppermint with a plastid targeting amino acid sequence, with or without a gene for biosynthesis of the precursor farnesyl diphosphate. Pure E?f was produced in stably transformed wheat lines with no other detectable phenotype but requiring targeting of the gene produced to the plastid. In laboratory behavioural assays, three species of cereal aphids were repelled and foraging was increased for a parasitic natural enemy. Although these studies show considerable potential for aphid control, field trials employing the single and double constructs showed no reduction in aphids or increase in parasitism. Insect numbers were low and climatic conditions erratic suggesting the need for further trials or a closer imitation, in the plant, of alarm pheromone release. PMID:26108150
Controlling malaria transmission with genetically-engineered, Plasmodium-resistant mosquitoes: milestones in a model system.
James, A A; Beerntsen, B T; Capurro, M de L; Coates, C J; Coleman, J; Jasinskiene, N; Krettli, A U
We are developing transgenic mosquitoes resistant to malaria parasites to test the hypothesis that genetically-engineered mosquitoes can be used to block the transmission of the parasites. We are developing and testing many of the necessary methodologies with the avian malaria parasite, Plasmodium gallinaceum, and its laboratory vector, Aedes aegypti, in anticipation of engaging the technical challenges presented by the malaria parasite, P. falciparum, and its major African vector, Anopheles gambiae. Transformation technology will be used to insert into the mosquito a synthetic gene for resistance to P. gallinaceum. The resistance gene will consist of a promoter of a mosquito gene controlling the expression of an effector protein that interferes with parasite development and/or infectivity. Mosquito genes whose promoter sequences are capable of sex- and tissue-specific expression of exogenous coding sequences have been identified, and stable transformation of the mosquito has been developed. We now are developing the expressed effector portion of the synthetic gene that will interfere with the transmission of the parasites. Mouse monoclonal antibodies that recognize the circumsporozoite protein of P. gallinaceum block sporozoite invasion of mosquito salivary glands, as well as abrogate the infectivity of sporozoites to a vertebrate host, the chicken, Gallus gallus, and block sporozoite invasion and development in susceptible cell lines in vitro. Using the genes encoding these antibodies, we propose to clone and express single-chain antibody constructs (scFv) that will serve as the effector portion of the gene that interferes with transmission of P. gallinaceum sporozoites. PMID:10697903
Oncolysis of Diffuse Hepatocellular Carcinoma by Intravascular Administration of a Replication-competent, Genetically Engineered Herpesvirus1
Timothy M. Pawlik; Hideo Nakamura; Sam S. Yoon; John T. Mullen; Soundararajalu Chandrasekhar; E. Antonio Chiocca; Kenneth K. Tanabe
Herpes simplex virus type 1 (HSV-1) replication within tumors can mediate tumor regression (oncolysis). The genetically engineered, HSV-1 mutant rRp450 does not express viral ribonucleotide reductase and is therefore replication conditional. During the course of infection, rRp450 expresses the cytochrome P450 transgene and HSV-1 thymidine kinase gene, thereby enabling it to bioactivate the prodrugs cyclophosphamide and ganciclovir, respectively. rRp450 replication
Effects of pH and acetic acid on glucose and xylose metabolism by a genetically engineered ethanologenic Escherichia coli
Hugh G. Lawford; Joyce D. Rousseau
Efficient utilization of the pentosan fraction of hemicellulose from lignocellulosic feedstocks offers an opportunity to increase\\u000a the yield and to reduce the cost of producing fuel ethanol. The patented, genetically engineered, ethanologenEscherichia coli B (pLOI297) exhibits high-performance characteristics with respect to both yield and productivity in xylose-rich lab media.\\u000a In addition to producing monomer sugar residues, thermochemical processing of biomass
Effects of pH and acetic acid on glucose and xylose metabolism by a genetically engineered ethanologenic Escherichia coli
H. G. Lawford; J. D. Rousseau
Efficient utilization of the pentosan fraction of hemicellulose from lignocellulosic feedstocks offers an opportunity to increase the yield and to reduce the cost of producing fuel ethanol. The patented, genetically engineered, ethanologen Escherichia coli B (pLOI297) exhibits high-performance characteristics with respect to both yield and productivity in xylose-rich lab media. In addition to producing monomer sugar residues, thermochemical processing of
In vivo delivery of recombinant human growth hormone from genetically engineered human fibroblasts implanted within Baxter immunoisolation devices
S. F. Josephs; T. Loudovaris; A. Dixit; S. K. Young; R. C. Johnson
Continuous delivery of therapeutic peptide to the systemic circulation would be the optimal treatment for a variety of diseases.\\u000a The Baxter TheraCyte? system is a membrane encapsulation system developed for implantation of tissues, cells such as endocrine cells or cell lines\\u000a genetically engineered for therapeutic peptide delivery in vivo. To demonstrate the utility of this system, cell lines were\\u000a developed
Ecological Risk Assessment of Alfalfa Medicago Varia L.) Genetically Engineered to Express a Human Metallothionein ( hMT ) Gene
Lidia S. Watrud; Santosh Misra; Leshitew Gedamu; Tamotsu Shiroyama; Sharon Maggard; George Di Giovanni
The objectives of these studies were two-fold: (1) to determine efficacy of low and high expression hMT gene constructs by assessing accumulation of Cu in shoots of parental and transgenic plants of alfalfa (Medicago varia L.) exposed to different concentrations of CuSO4 by addition of CuSO4 solutions to soil and (2) to identify potential unintended effects of the genetic engineering
Recommendations for the design of laboratory studies on non-target arthropods for risk assessment of genetically engineered plants
J÷rg Romeis; Richard L. Hellmich; Marco P. Candolfi; Keri Carstens; Adinda De Schrijver; Angharad M. R. Gatehouse; Rod A. Herman; Joseph E. Huesing; Morven A. McLean; Alan Raybould; Anthony M. Shelton; Annabel Waggoner
This paper provides recommendations on experimental design for early-tier laboratory studies used in risk assessments to evaluate\\u000a potential adverse impacts of arthropod-resistant genetically engineered (GE) plants on non-target arthropods (NTAs). While\\u000a we rely heavily on the currently used proteins from Bacillus thuringiensis (Bt) in this discussion, the concepts apply to other arthropod-active proteins. A risk may exist if the newly
Automatic Analogue Network Synthesis using Genetic Algorithms IEE/IEEE International Conference on Genetic Algorithms in Engineering Systems: Innovations and
Grimbleby, James
that GAs are both practical and robust. Application of GAs to Network Synthesis Each individualAutomatic Analogue Network Synthesis using Genetic Algorithms IEE/IEEE International Conference provide a basis for automatic synthesis of analogue electronic networks. Passive linear networks have been
Our research objectives were to: (1) determine the persistence of an introduced surrogate (Cellulomonas sp NRC 2406) for a genetically engineered microorganism (GEM) in three streamlined habitats; sediments, growths of Cladophora (Chlorophyta), and leaf packs, (2) test ommunity a...
Reverse-engineering the genetic circuitry of a cancer cell with predicted intervention in chronic lymphocytic leukemia
Vallat, Laurent; Kemper, Corey A.; Jung, Nicolas; Maumy-Bertrand, Myriam; Bertrand, Frķdķric; Meyer, Nicolas; Pocheville, Arnaud; Fisher, John W.; Gribben, John G.; Bahram, Seiamak
Cellular behavior is sustained by genetic programs that are progressively disrupted in pathological conditionsŚnotably, cancer. High-throughput gene expression profiling has been used to infer statistical models describing these cellular programs, and development is now needed to guide orientated modulation of these systems. Here we develop a regression-based model to reverse-engineer a temporal genetic program, based on relevant patterns of gene expression after cell stimulation. This method integrates the temporal dimension of biological rewiring of genetic programs and enables the prediction of the effect of targeted gene disruption at the system level. We tested the performance accuracy of this model on synthetic data before reverse-engineering the response of primary cancer cells to a proliferative (protumorigenic) stimulation in a multistate leukemia biological model (i.e., chronic lymphocytic leukemia). To validate the ability of our method to predict the effects of gene modulation on the global program, we performed an intervention experiment on a targeted gene. Comparison of the predicted and observed gene expression changes demonstrates the possibility of predicting the effects of a perturbation in a gene regulatory network, a first step toward an orientated intervention in a cancer cell genetic program. PMID:23267079
Modified H5 promoter improves stability of insert genes while maintaining immunogenicity during extended passage of genetically engineered MVA vaccines.
Wang, Zhongde; Martinez, Joy; Zhou, Wendi; La Rosa, Corinna; Srivastava, Tumul; Dasgupta, Anindya; Rawal, Ravindra; Li, Zhongqui; Britt, William J; Diamond, Don
We have engineered recombinant (r) Modified Vaccinia Ankara (MVA) to express multiple antigens under the control of either of two related vaccinia synthetic promoters (pSyn) with early and late transcriptional activity or the modified H5 (mH5) promoter which has predominant early activity. We sequentially passaged these constructs and analyzed their genetic stability by qPCR, and concluded that rMVA expressing multiple antigens using the mH5 promoter exhibit remarkable genetic stability and maintain potent immunogenicity after serial passage. In contrast, rMVA expressing antigens using engineered vaccinia synthetic E/L (pSyn I or II) promoters are genetically unstable. Progressive accumulation of antigen loss variants resulted in a viral preparation with lower immunogenicity after serial passage. Metabolic labeling, followed by cold chase revealed little difference in stability of proteins expressed from mH5 or pSyn promoter constructs. We conclude that maintenance of genetic stability which is achieved using mH5, though not with pSyn promoters, is linked to timing, not the magnitude of expression levels of foreign antigen, which is more closely associated with immunogenicity of the vaccine. PMID:19969118
Can Man Control His Biological Evolution? A Symposium on Genetic Engineering. Man's Responsibility to His Future
ERIC Educational Resources Information Center
Hoagland, Hudson
Biological evolution can be carried out in the laboratory. With new knowledge available in genetics, possibilities are raised that genetic characters can be transferred in the future to embryos according to a predetermined plan. (PS)
Engineered sensors and genetic regulatory networks for control of cellular metabolism
Moser, Felix, Ph. D. Massachusetts Institute of Technology
Complex synthetic genetic programs promise unprecedented control over cellular metabolism and behavior. In this thesis, I describe the design and development of a synthetic genetic program to detect conditions underlying ...
NSDL National Science Digital Library
The Tech Museum of Innovation
This online tutorial from the TheTech Museum of Innovation focuses on genetics. The interactive topics will initially introduce the user to the DNA, chromosomes, and the make up of human genes. Further topics will examine forensic science, the history of forensics, fingerprinting, and cloning background research and community response to cloning. Finally, the resource provides connections to gallery exhibits, science labs, and a design challenge that engages the learner to write a persuasive letter to a group or organization responsible for cloning or DNA decision making. Copyright 2005 International Technology Education Association
Implicit axioms - a main paradigm within the discourse between science and religion and its relevance for the debate on genetic engineering as well as artificial intelligence
Astrid Dinter
It can be shown that model-building in genetics is already connected with implicit axioms which partially are connected with metaphysical implications. This is especially true for human genetic engineering. In this context, a dialogue with theology could be very fruitful because these implicit axioms can be analyzed and the proposed models of humankind reworked. It will be very important for
Oxidative stress and physiological, epigenetic and genetic variability in plant tissue culture: implications for micropropagators and genetic engineers
Alan C. Cassells; Rosario F. Curry
A number of well defined problems in physiological, epigenetic and genetic quality are associated with the culture of plant\\u000a cell, tissue and organs in vitro, namely, absence or loss of organogenic potential (recalcitrance), hyperhydricity (`vitrification') and somaclonal variation.\\u000a These broad terms are used to describe complex phenomena that are known to be genotype and environment dependent. These phenomena\\u000a affect the
Animal models of adrenocortical tumorigenesis
Beuschlein, Felix; Galac, Sara; Wilson, David B.
Over the past decade, research on human adrenocortical neoplasia has been dominated by gene expression profiling of tumor specimens and by analysis of genetic disorders associated with a predisposition to these tumors. Although these studies have identified key genes and associated signaling pathways that are dysregulated in adrenocortical neoplasms, the molecular events accounting for the frequent occurrence of benign tumors and low rate of malignant transformation remain unknown. Moreover, the prognosis for patients with adrenocortical carcinoma remains poor, so new medical treatments are needed. Naturally occurring and genetically engineered animal models afford a means to investigate adrenocortical tumorigenesis and to develop novel therapeutics. This comparative review highlights adrenocortical tumor models useful for either mechanistic studies or preclinical testing. Three model species ¢ mouse, ferret, and dog ¢ are reviewed, and their relevance to adrenocortical tumors in humans is discussed. PMID:22100615
Modeling Sparse Engine Test Data Using Genetic Programming Chevron Information Technology Company
Fernandez, Thomas
-world meaning. We hope the results of this study would benefit other engine oil modeling applications. 1 INTRODUCTION Laboratory engine tests are among the tools used to measure engine oil performance. These tests are specified in various engine oil performance categories for licensing and certification (API, 1999; ASTM 1999
Genetically Engineered Immunomodulatory Streptococcus thermophilus Strains Producing Antioxidant Enzymes Exhibit Enhanced Anti-Inflammatory Activities
del Carmen, Silvina; de Moreno de LeBlanc, Alejandra; Martin, Rebeca; Chain, Florian; Langella, Philippe; Berm·dez-Humarßn, Luis G.
The aims of this study were to develop strains of lactic acid bacteria (LAB) having both immunomodulatory and antioxidant properties and to evaluate their anti-inflammatory effects both in vitro, in different cellular models, and in vivo, in a mouse model of colitis. Different Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains were cocultured with primary cultures of mononuclear cells. Analysis of the pro- and anti-inflammatory cytokines secreted by these cells after coincubation with candidate bacteria revealed that L. delbrueckii subsp. bulgaricus CRL 864 and S. thermophilus CRL 807 display the highest anti-inflammatory profiles in vitro. Moreover, these results were confirmed in vivo by the determination of the cytokine profiles in large intestine samples of mice fed with these strains. S. thermophilus CRL 807 was then transformed with two different plasmids harboring the genes encoding catalase (CAT) or superoxide dismutase (SOD) antioxidant enzymes, and the anti-inflammatory effects of recombinant streptococci were evaluated in a mouse model of colitis induced by trinitrobenzenesulfonic acid (TNBS). Our results showed a decrease in weight loss, lower liver microbial translocation, lower macroscopic and microscopic damage scores, and modulation of the cytokine production in the large intestines of mice treated with either CAT- or SOD-producing streptococci compared to those in mice treated with the wild-type strain or control mice without any treatment. Furthermore, the greatest anti-inflammatory activity was observed in mice receiving a mixture of both CAT- and SOD-producing streptococci. The addition of L. delbrueckii subsp. bulgaricus CRL 864 to this mixture did not improve their beneficial effects. These findings show that genetically engineering a candidate bacterium (e.g., S. thermophilus CRL 807) with intrinsic immunomodulatory properties by introducing a gene expressing an antioxidant enzyme enhances its anti-inflammatory activities. PMID:24242245
The potential of transcription factor-based genetic engineering in improving crop tolerance to drought.
Rabara, Roel C; Tripathi, Prateek; Rushton, Paul J
Drought is one of the major constraints in crop production and has an effect on a global scale. In order to improve crop production, it is necessary to understand how plants respond to stress. A good understanding of regulatory mechanisms involved in plant responses during drought will enable researchers to explore and manipulate key regulatory points in order to enhance stress tolerance in crops. Transcription factors (TFs) have played an important role in crop improvement from the dawn of agriculture. TFs are therefore good candidates for genetic engineering to improve crop tolerance to drought because of their role as master regulators of clusters of genes. Many families of TFs, such as CCAAT, homeodomain, bHLH, NAC, AP2/ERF, bZIP, and WRKY have members that may have the potential to be tools for improving crop tolerance to drought. In this review, the roles of TFs as tools to improve drought tolerance in crops are discussed. The review also focuses on current strategies in the use of TFs, with emphasis on several major TF families in improving drought tolerance of major crops. Finally, many promising transgenic lines that may have improved drought responses have been poorly characterized and consequently their usefulness in the field is uncertain. New advances in high-throughput phenotyping, both greenhouse and field based, should facilitate improved phenomics of transgenic lines. Systems biology approaches should then define the underlying changes that result in higher yields under water stress conditions. These new technologies should help show whether manipulating TFs can have effects on yield under field conditions. PMID:25118806
[Atrazine wastewater treatment in a SPG membrane-aerated genetically engineered microorganism biofilm reactor].
Liu, Chun; Gong, Peng-Fei; Xiao, Tai-Min; Zhang, Ming; Nian, Yong-Jia; Yang, Jing-Liang; Zhang, Jing
Membrane-aerated biofilm reactor (MABR) represent a novel membrane-biological wastewater treatment technology. In addition, bioaugmented treatment using genetically engineered microorganism (GEM) biofilm in MABR is proposed to improve refractory pollutant removal. In the present study, a SPG membrane aerated-biofilm reactor (SPG-MABR) with GEM biofilm formed on the SPG membrane surface was applied to treat atrazine wastewater. The influences of air pressure, biofilm biomass and liquid velocity on the performance of the SPG-MABR were investigated. The variation of GEM biofilm during the SPG-MABR operation was observed. The results indicated that the increased air pressure could promote atrazine and COD removal as well as re-oxygenation by increasing oxygen permeability coefficient. A higher biofilm biomass could also enhance atrazine and COD removal, but simultaneously reduce the re-oxygenation rate because biofilm thickness and oxygen transfer resistance increased. When liquid velocity in the SPG-MABR was decreased under laminar flow condition, atrazine and COD removal was improved due to the facilitated contaminant diffusion from wastewater to biofilm. The atrazine removal efficiency reached to 98.6% in the SPG-MABR after 5d treatment at air pressure of 300 kPa, biofilm biomass of 25 g x m(-2) and liquid velocity of 0.05 m x s(-1). The microbial polymorphism of GEM biofilm was observed during the SPG-MABR operation. The surface of GEM biofilm was gradually covered by other microbial cells and the distribution of GEM cells reduced, but inside the GEM biofilm, the GEM cells were still dominant. PMID:25338374
Production of human lactoferrin and lysozyme in the milk of transgenic dairy animals: past, present, and future.
Cooper, Caitlin A; Maga, Elizabeth A; Murray, James D
Genetic engineering, which was first developed in the 1980s, allows for specific additions to animals' genomes that are not possible through conventional breeding. Using genetic engineering to improve agricultural animals was first suggested when the technology was in the early stages of development by Palmiter et al. (Nature 300:611-615, 1982). One of the first agricultural applications identified was generating transgenic dairy animals that could produce altered or novel proteins in their milk. Human milk contains high levels of antimicrobial proteins that are found in low concentrations in the milk of ruminants, including the antimicrobial proteins lactoferrin and lysozyme. Lactoferrin and lysozyme are both part of the innate immune system and are secreted in tears, mucus, and throughout the gastrointestinal (GI) tract. Due to their antimicrobial properties and abundance in human milk, multiple lines of transgenic dairy animals that produce either human lactoferrin or human lysozyme have been developed. The focus of this review is to catalogue the different lines of genetically engineered dairy animals that produce either recombinant lactoferrin or lysozyme that have been generated over the years as well as compare the wealth of research that has been done on the in vitro and in vivo effects of the milk they produce. While recent advances including the development of CRISPRs and TALENs have removed many of the technical barriers to predictable and efficient genetic engineering in agricultural species, there are still many political and regulatory hurdles before genetic engineering can be used in agriculture. It is important to consider the substantial amount of work that has been done thus far on well established lines of genetically engineered animals evaluating both the animals themselves and the products they yield to identify the most effective path forward for future research and acceptance of this technology. PMID:26059245
Gabapentin decreases the severity of dystonia at low doses in a genetic animal model of paroxysmal dystonic choreoathetosis.
Richter, A; L÷scher, W
The effects of the gamma-aminobutyric acid (GABA)-potentiating drug gabapentin (1-(aminomethyl) cyclohexaneacetic acid) on severity of dystonia were examined in a hamster model of idiopathic paroxysmal dystonic choreoathetosis. In the genetically dystonic hamster (dt(sz)) recent pharmacological and neurochemical studies suggested that disturbed GABAergic inhibition is involved in the pathogenesis. In line with a case report of beneficial effects in human paroxysmal dystonic choreoathetosis, gabapentin reduced the severity of dystonia in mutant hamsters at doses of 5 and 10 mg kg(-1) i.p. At higher doses (20 and 100 mg kg(-1)), gabapentin, however, failed to exert antidystonic effects. The GABApotentiating activity of gabapentin could explain the antidystonic effects of low doses, while the loss of efficacy at higher doses may be due to other mechanisms of gabapentin. PMID:10225372
Prediction of genetic values of sires for growth traits of crossbred cattle using a multivariate animal model with heterogeneous variances.
N·±ez-Dominguez, R; Van Vleck, L D; Cundiff, L V
The purpose of this study was to evaluate the effect of adjusting for heterogeneous variances across breed groups on prediction of breeding values (PBV) of selected sires and on breed of sire effects. Data on weights at birth (BWT), 200 d (WW), and 365 d (YW) of purebred and crossbred calves from matings of Angus (A), Hereford (H), Polled Hereford, Charolais, Shorthorn, Simmental, Limousin, Maine-Anjou, Chianina, Gelbvieh, Tarentaise, and Salers Bulls to A and H cows were used. Calf performance in H and A dams was treated as a different trait. Models compared included fixed birth year, cow age, and sex classes and crossbreeding effect as a covariate; random direct and maternal genetic and permanent environmental effects were also included, but their variance structure was different. Model I assumed homogeneous variances across breed groups. Model II accounted for heterogeneous variances. Sires were ranked based on PBV from each model, and means of PBV of selected sires were calculated based on Model II. Differences between mean PBV were small for BWT, intermediate for WW, and larger for YW. Differences in PBV of selected sires increased as selection intensity increased, but only for WW and YW. Large differences in mean PBV of selected sires between maternal environments (H vs A) were observed for WW and YW for various sire breeds. Means of PBV of selected sires based on Model II exceeded those based on Model I by 6 to 16 kg of YW for various selection intensities and maternal environments. Estimates of breed of sire effects from Model I or II were similar for BWT and WW, but large differences were found for YW. Results indicate that some additional economic returns may be gained by commercial producers if sires are chosen across breeds based on predicted genetic values computed with models accounting for heterogeneous variances. PMID:8617664
NSDL National Science Digital Library
Understanding Animal Research (Understanding Animal Research)
The public debate on animal research sometimes gets so heated that the facts can be overlooked. How many animals are used in research every year? Do people know that most of them are mice or rats? Why are animals genetically modified? How is animal research regulated? How are the animals cared for? What actually happens to research animals? How does the use of animals in research and testing compare with other uses of animals by society? This website aims to answer all of these questions as well as provide information on animal research and human health, policy issues, and latest news. This website also includes a learning center. Information is geared towards learners in the U.K.
An improved ARS2-derived nuclear reporter enhances the efficiency and ease of genetic engineering in Chlamydomonas.
Specht, Elizabeth A; Nour-Eldin, Hussam Hassan; Hoang, Kevin T D; Mayfield, Stephen P
The model alga Chlamydomonas reinhardtii has been used to pioneer genetic engineering techniques for high-value protein and biofuel production from algae. To date, most studies of transgenic Chlamydomonas have utilized the chloroplast genome due to its ease of engineering, with a sizeable suite of reporters and well-characterized expression constructs. The advanced manipulation of algal nuclear genomes has been hampered by limited strong expression cassettes, and a lack of high-throughput reporters. We have improved upon an endogenous reporter gene - the ARS2 gene encoding an arylsulfatase enzyme - that was first cloned and characterized decades ago but has not been used extensively. The new construct, derived from ARS2 cDNA, expresses significantly higher levels of reporter protein and transforms more efficiently, allowing qualitative and quantitative screening using a rapid, inexpensive 96-well assay. The improved arylsulfatase expression cassette was used to screen a new transgene promoter from the ARG7 gene, and found that the ARG7 promoter can express the ARS2 reporter as strongly as the HSP70-RBCS2 chimeric promoter that currently ranks as the best available promoter, thus adding to the list of useful nuclear promoters. This enhanced arylsulfatase reporter construct improves the efficiency and ease of genetic engineering within the Chlamydomonas nuclear genome, with potential application to other algal strains. PMID:25224580
Genetically engineered, live, attenuated vaccines protect nonhuman primates against aerosol challenge with a virulent IE strain of Venezuelan equine encephalitis virus.
Reed, Douglas S; Lind, Cathleen M; Lackemeyer, Matthew G; Sullivan, Lawrence J; Pratt, William D; Parker, Michael D
Two live, attenuated strains of Venezuelan equine encephalitis virus (VEE), IE1150K and V3526, were administered to macaques to determine if they could elicit protection against an aerosol challenge with virulent VEE virus of the IE variety (VEEV-IE). These viruses were rescued from full-length cDNA clones of 68U201 (VEEV-IE variety) and Trinidad donkey (VEEV-IA/B variety), respectively, and both have a furin cleavage site deletion mutation and a second-site resuscitating mutation. Both vaccines elicited neutralizing antibodies to viruses of the homologous variety but not to viruses of the heterologous variety. Eight weeks after vaccination, the macaques were challenged by aerosol exposure to virulent 68U201. Macaques vaccinated with V3526 were protected as well as macaques inoculated with IE1009, the wild-type infectious clone of 68U201. However, IE1150K failed to significantly protect macaques relative to controls. V3526 has now been shown to protect macaques against both IA/B [Pratt WD, Davis NL, Johnston RE, Smith JF. Genetically engineered, live attenuated vaccines for Venezuelan equine encephalitis: testing in animal models. Vaccine 2003;21(25-26):3854-62] and IE strains of VEE viruses. PMID:15837213
The potential of genetic engineering for improving brewing, wine-making and baking yeasts
S. Dequin
The end of the twentieth century was marked by major advances in life technology, particularly in areas related to genetics and more recently genomics. Considerable progress was made in the development of genetically improved yeast strains for the wine, brewing and baking industries. In the last decade, recombinant DNA technology widened the possibilities for introducing new properties. The most remarkable
GENETIC ENGINEERING - GENETECHNOLOGY IS IT SALVATION OR CURSE FOR THE 21 th CENTURY?
Arpad Pusztai
With progress in genetic science it became possible for the first time to transfer genetic information derived from various sources in to microorganisms. With this major advance in the seventies of the last century a new scientific research field has opened up, what we now call as the recombinant DNA technology. By taking a gene from any source and transferring
Harvard U.'s Request for Commercial Rights to New Strain of Mouse Forces Debate in Europe over Whether Animals Can Be Patented.
ERIC Educational Resources Information Center
Chronicle of Higher Education, 1989
The European Patent Convention has informed Harvard University that its application for a patent on a genetically engineered mouse may be refused. The application was the first to obtain patent protection across most of Europe for a transgenic animal, one which has been implanted with genes from another animal. (MSE)
Genetic Diversity and Antimicrobial Resistance of Escherichia coli from Human and Animal Sources Uncovers Multiple Resistances from Human Sources
Ibekwe, A. Mark; Murinda, Shelton E.; Graves, Alexandria K.
Escherichia coli are widely used as indicators of fecal contamination, and in some cases to identify host sources of fecal contamination in surface water. Prevalence, genetic diversity and antimicrobial susceptibility were determined for 600 generic E. coli isolates obtained from surface water and sediment from creeks and channels along the middle Santa Ana River (MSAR) watershed of southern California, USA, after a 12 month study. Evaluation of E. coli populations along the creeks and channels showed that E. coli were more prevalent in sediment compared to surface water. E. coli populations were not significantly different (P?=?0.05) between urban runoff sources and agricultural sources, however, E. coli genotypes determined by pulsed-field gel electrophoresis (PFGE) were less diverse in the agricultural sources than in urban runoff sources. PFGE also showed that E. coli populations in surface water were more diverse than in the sediment, suggesting isolates in sediment may be dominated by clonal populations.Twenty four percent (144 isolates) of the 600 isolates exhibited resistance to more than one antimicrobial agent. Most multiple resistances were associated with inputs from urban runoff and involved the antimicrobials rifampicin, tetracycline, and erythromycin. The occurrence of a greater number of E. coli with multiple antibiotic resistances from urban runoff sources than agricultural sources in this watershed provides useful evidence in planning strategies for water quality management and public health protection. PMID:21687635
Artificial Animals for Computer Animation
Toronto, University of
animals. We create self-animating, autonomous agents which emulate the realistic appearance, movementArtificial Animals for Computer Animation: Biomechanics, Locomotion, Perception, and Behavior Ī Xiaoyuan Tu 1996 ALL RIGHTS RESERVED #12;Artificial Animals for Computer Animation: Biomechanics
AnimAl BehAvior AnimAl Biology Anthropology AviAn ScienceS BiochemiStry, moleculAr, cellulAr AnD DevelopmentAl Biology BiologicAl SyStemS engineering BiomeDicAl engineering BiophySicS BioStAtiSticS clinicAl reSeArch
Schladow, S. Geoffrey
, human evolution, molecular anthropology, archaeological theory, hunter-gatherers, and North American, animal management and welfare, physiology and behavior, behavior and conservation, primate behavior, behavioral ecology of nonhuman primates, biological anthropology, evolution of primate and human behavior
Genetically engineered cardiac pacemaker: Stem cells transfected with HCN2 gene and myocytesŚA model
NASA Astrophysics Data System (ADS)
Kanani, S.; Pumir, A.; Krinsky, V.
One of the successfully tested methods to design genetically engineered cardiac pacemaker cells consists in transfecting a human mesenchymal stem cell (hMSC) with a HCN2 gene and connecting it to a myocyte. We develop and study a mathematical model, describing a myocyte connected to a hMSC transfected with a HCN2 gene. The cardiac action potential is described both with the simple Beeler Reuter model, as well as with the elaborate dynamic Luo Rudy model. The HCN2 channel is described by fitting electrophysiological records, in the spirit of Hodgkin Huxley. The model shows that oscillations can occur in a pair myocyte-stem cell, that was not observed in the experiments yet. The model predicted that: (1) HCN pacemaker channels can induce oscillations only if the number of expressed I channels is low enough. At too high an expression level of I channels, oscillations cannot be induced, no matter how many pacemaker channels are expressed. (2) At low expression levels of I channels, a large domain of values in the parameter space (n, N) exists, where oscillations should be observed. We denote N the number of expressed pacemaker channels in the stem cell, and n the number of gap junction channels coupling the stem cell and the myocyte. (3) The expression levels of I channels observed in ventricular myocytes, both in the Beeler Reuter and in the dynamic Luo Rudy models are too high to allow to observe oscillations. With expression levels below ś1/4 of the original value, oscillations can be observed. The main consequence of this work is that in order to obtain oscillations in an experiment with a myocyte-stem cell pair, increasing the values of n, N is unlikely to be helpful, unless the expression level of I has been reduced enough. The model also allows us to explore levels of gene expression not yet achieved in experiments, and could be useful to plan new experiments, aimed at improving the robustness of the oscillations.
Biomimetic Fabrication of Genetically-Engineered Collagen Peptide-Assembled Freestanding Films Reinforced by Quantum Dot Joints.
Wei, Zengyan; Maeda, Yoshiaki; Matsui, Hiroshi
Genetically-engineered collagen peptides were assembled into freestanding films when QDs are co-assembled as joints between collagen domains. These peptide based films show excellent mechanical properties with Young's modulus of ~20 GPa, much larger than most of multi-composite polymer films and previously reported freestanding nanoparticle-assembled sheets, and it is even close to the bone tissue in nature. These films show little permanent deformation under small indentation while the mechanical hysteresis becomes remarkable when the load approaches near and beyond the rupture point, which is also characteristic to the bone tissue. PMID:22982983
Genetic engineering combined with deep UV resonance Raman spectroscopy for structural characterization of amyloid-like fibrils.
Sikirzhytski, Vitali; Topilina, Natalya I; Higashiya, Seiichiro; Welch, John T; Lednev, Igor K
Elucidating the structure of the cross-beta core in large amyloid fibrils is a challenging problem in modern structural biology. For the first time, a set of de novo polypeptides was genetically engineered to form amyloid-like fibrils with similar morphology and yet different strand length. Differential ultraviolet Raman spectroscopy allowed for separation of the spectroscopic signatures of the highly ordered beta-sheet strands and turns of the fibril core. The relationship between Raman frequencies and Ramachandran dihedral angles of the polypeptide backbone indicates the nature of the beta-sheet and turn structural elements. PMID:18410104
Imipramine treatment increases the number of hippocampal synapses and neurons in a genetic animal model of depression.
Chen, Fenghua; Madsen, Torsten M; Wegener, Gregers; Nyengaard, Jens R
The aim was to investigate treatment effects of the antidepressant imipramine on the markers of neuronal plasticity. We investigated changes in neuron and synapse numbers in a rat strain that displays a genetic susceptibility to depressive behavior, the Flinders Sensitive and Resistant Lines (FSL/FRL). All rats were treated with imipramine (15 mg/kg) or saline (i.p) once daily for 25 days. The volume, neuron and synapse numbers in the hippocampus were estimated using design-based stereological methods. Under untreated conditions, the volume and the number of neurons and synapses were significantly smaller in the FSL saline group (untreated "depressed" rats) compared with the FRL saline group (normal rats), showing correlation to the observed decreased immobility in the forced swim test. Imipramine treatment significantly increased the number of neurons in the granule cell layer (GCL) and spine synapses in the CA1 in the FSL imipramine group (treated "depressed" rats) compared with the FSL saline group. The neuron numbers in the GCL and Hilus showed no differences in the FSL imipramine group compared to the FRL saline group. In conclusion, baseline levels of the volume and the number of neurons and spine synapses in hippocampus were significantly smaller in the untreated FSL rats. Our findings indicate that chronic imipramine treatment reverses the suppression of neurogenesis and synaptogenesis in the hippocampus of the "depressed" FSL rats, and this occurs in correlation with behavioral effects. Our results support the neuronal plasticity hypothesis that depressive disorders may be related to impairments of structural plasticity and neuronal viability in hippocampus, furthermore, antidepressant treatment counteracts the structural impairments. PMID:19921703
Genetic diversity of Escherichia coli isolates of animal and environmental origins from an integrated poultry production chain.
Pasquali, Frķdķrique; Lucchi, Alex; Braggio, Simonetta; Giovanardi, Davide; Franchini, Achille; Stonfer, Maurizio; Manfreda, Gerardo
Escherichia coli is a normal inhabitant of the intestinal tract of chickens, but when an imbalance in the gut microbiota occurs, E. coli may overgrow and cause extraintestinal infections. The aims of this study were to assess the distribution and spread of E. coli isolates with specific phylogenetic groups and antimicrobial resistance characters among asymptomatic breeder flocks and their broiler progenies with early symptoms of colibacillosis. Broiler flocks were treated with lincospectin during the first week of life and sampled at one, 21 and 42 days. The majority of the 363 E. coli isolates belonged to phylogenetic group A (53.17%), followed by groups D (23.14%), B1 (19.28%) and B2 (4.41%). In broilers, group A was the most represented in birds of 21 and 42 days of age whereas group B1 was the most represented phylogroup in one-day old chicks. More than 90.00% of the isolates were resistant to one or more antimicrobials. Along the life-time of broilers, no differences were found on the occurrence of resistant isolates except for the number of E. coli with elevated MIC to spectinomycin, which increased significantly after the lincospectin treatment. According to XbaI-macrorestriction analysis, a high genetic diversity among E. coli isolates was underlined. Four antimicrobial resistant E. coli isolates of phylogroups A, B1 and D collected from breeders showed similar PFGE patterns to five isolates collected from the respective broiler progenies suggesting a potential spread of these isolates from breeders to broilers. PMID:26013418
Bioterrorism: intentional introduction of animal disease.
Clarke, N P; Rinderknecht, J L
The possibility of the intentional introduction of animal disease as an act of bioterrorism adds a new dimension to the development of strategies for assessment, prevention, response and recovery from exotic diseases, including the zoonoses. The vulnerability of livestock operations, the likelihood of success, the possibility of the use of genetically engineered organisms and limited resources to handle multiple outbreaks place new pressures on policy-makers and emergency responders to make best use of limited resources. The methods for managing a natural occurrence or accidental introduction of high-consequence diseases are generally applicable to containment and recovery from outbreaks of intentionally introduced animal diseases. Zoonotic agents increase the complexity at both international and national levels. Modern biology provides both increased threat of new disease entities and methods for earlier and more effective detection and intervention. Improved methods are emerging for defining trade restrictions and animal movement and for determining when it is safe to resume normal trade. PMID:21809759
Genetic Engineering of Beta-Carotene Production in Honeydew Melons (Cucumis melo L. inodorus)Ā
Ren, Yan
Genetic transformation is a useful tool to incorporate novel genes, potentially allowing sexual incompatibility and interspecific barriers to be circumvented. The purpose of this study was to improve beta-carotene levels ...
Engineered ascorbate peroxidase as a genetically encoded reporter for electron microscopy
Deerinck, Thomas J
Electron microscopy (EM) is the standard method for imaging cellular structures with nanometer resolution, but existing genetic tags are inactive in most cellular compartments[superscript 1] or require light and can be ...
Can Man Control His Biological Evolution? A Symposium on Genetic Engineering. Xeroxing Human Beings
ERIC Educational Resources Information Center
Freund, Paul A.
If the aim of new research is to improve the genetic inheritance of future generations, then decisions regarding who should decide what research should be done needs to be established. Positive and negative eugenics need to be considered thoroughly. (PS)
Towards the development of better crops by genetic transformation using engineered plant chromosomes
Manoj K. DharSanjana; Sanjana Kaul; Jasmeet Kour
Plant Biotechnology involves manipulation of genetic material to develop better crops. Keeping in view the challenges being\\u000a faced by humanity in terms of shortage of food and other resources, we need to continuously upgrade the genomic technologies\\u000a and fine tune the existing methods. For efficient genetic transformation, Agrobacterium-mediated as well as direct delivery methods have been used successfully. However, these
Genetically enhanced cows resist intramammary Staphylococcus aureus infection
Anne M Powell; Max J Paape; David E Kerr; Douglas D Bannerman; Vernon G Pursel; Kevin D Wells; Neil Talbot; Harold W Hawk; Robert J Wall
Mastitis, the most consequential disease in dairy cattle, costs the US dairy industry billions of dollars annually. To test the feasibility of protecting animals through genetic engineering, transgenic cows secreting lysostaphin at concentrations ranging from 0.9 to 14 mg\\/ml in their milk were produced. In vitro assays demonstrated the milk's ability to kill Staphylococcus aureus. Intramammary infusions of S. aureus
Animal cloning: problems and prospects
D. N. Wells
Summary An efficient animal cloning technology would provide many new opportunities for livestock agriculture, human medicine, and animal conservation. Nuclear cloning involves the production of animals that are genetically identical to the donor cells used in a technique known as nuclear transfer (NT). However, at present it is an inefficient process: in cattle, only around 6% of the embryos transferred
Since the early 1970's, plant biotechnology research and development has progressed dramatically. enetically engineered plants (GEPS) that express new or improved pesticidal properties, enhanced resistance to herbicides, improved nutritional value, and resistance to disease have ...
Genetically engineered maize plants reveal distinct costs and benefits of constitutive volatile emissions in the field
Technology Transfer Automated Retrieval System (TEKTRAN)
Genetic manipulation of plant volatile emissions is a promising tool to enhance plant defences against herbivores. However, the potential costs associated with the manipulation of specific volatile synthase genes are unknown. Therefore, we investigated the physiological and ecological effects of tra...
Gene Flow in Genetically Engineered Perennial Grasses: Lessons for Modification of Dedicated Bioenergy Crops
Technology Transfer Automated Retrieval System (TEKTRAN)
Genetic modification of dedicated bioenergy crops, such as switchgrass, will play a major role in crop improvement for a wide range of beneficial traits specific to biofuels. One obstacle that arises regarding transgenic improvement of perennials used for biofuels is the propensity of these plants t...
Cancer stem cells with genetic instability: the best vehicle with the best engine for cancer
E Lagasse
Our understanding of the role of stem cells in cancer development is evolving quickly. In the course of tumor expansion, a subpopulation of tumor cells with stem cell-like features has been noted. These cancer stem cells give rise to transit amplifying tumor cells, which comprise the majority of the tumor mass prior to terminal differentiation. Combining this finding with genetic
Genetic engineering within the adult brain: Implications for molecular approaches to behavioral neuroscience
Pedro R. Lowenstein; Maria G. Castro
Currently, the most popular technology used to modify the molecular makeup of the nervous system is through germline modifications of early embryos. This allows to construct gene æknock-insÆ (gene overexpression) or æknock-outsÆ (gene deletions). This technology leads to gene additions or deletions from the earliest developmental stages. This can potentially lead to compensatory genetic changes. The technology to achieve inducible
Environmental Risk Assessment of Genetically Engineered Herbicide-Tolerant Zoysia japonica
T. W. Bae; E. Vanjildorj; S. Y. Song; S. Nishiguchi; Yang Chonnam; I. J. Song; T. Chandrasekhar; T. W. Kang; Kim Chonnam; J. Koh; S. Y. Park; J. Lee; K. H. Ryu; K. Z. Riu; P.-S. Song; H. Y. Lee
Herbicide-tolerant Zoysia grass (Zoysia japonica Steud.) has been generated previously through Agrobacterium tumefaciens- mediated transformation. Th e genetically modifi ed (GM) Zoysia grass survived Basta spraying and grew to maturity normally while the wild-type (WT) grass stopped growing and died. GM Zoysia grass will permit more effi cient weed control for various turf grass plantings such as home lawns, golf
NSDL National Science Digital Library
This animation, created by faculty at Dartmouth University, is a boost converter. The resource features other animations such as buck and discontinuous converters. They also add simple diode, bridge and half-wave rectifiers. Although simple in design, this can still be a useful resource for those interested in electrical engineering.
J. Nielsen
Metabolic engineering has developed as a very powerful approach to optimising industrial fermentation processes through the introduction of directed genetic changes using recombinant DNA technology. Successful metabolic engineering starts with a careful analysis of cellular function; based on the results of this analysis, an improved strain is designed and subsequently constructed by genetic engineering. In recent years some very powerful
[Experimental animal research in the EU legislation].
Bolliger, Gieri
The legislation of the European Union captures experimental animal research only in part. Binding standards can be found in various legal records of the community-legislation, and in particular in the so-called guideline for animal experimentation 86/609/EWG. But these guidelines do not represent an actual animal protection measure but rather one of harmonisation with the primary goal of unification of the regulations of the participating states in order to prevent distortions of competition and trade barriers which could harm the common market. Although the guideline contains some practicable approaches in the direction of an up-to-date animal experimentation law, it only defines general goals which allows for considerable leeway in national implementation within the individual EU countries, and it has only a limited area of legal operation. On the one hand, only vertebrae are being included, and on the other, the law is only being applied in the area of applied research and protects only animals used in product- and substance-development or test procedures as well as those used in the framework of environmental protection. Various important fields of research are thus not subject to a common regulation and are assigned to national regulation. This concerns animal experimentation in education and training or for military or so-called defence-relevant medical purposes and, in particular, the whole area of basic research including the field of genetic engineering in animals with it's growing significance. The guideline is in need of widening it's scope of application as well as of various adjustments to recent scientific findings and developments in order to become suitable as a more restrictive animal protection law on the community level. It could also be desirable to include animal protection into the catalogue of community-goals in order to make it an independent component of the politics of the Union and to establish the groundwork for a decree of comprehensive and efficient regulations. PMID:12096325
Quantification of Tumor Burden in a Genetically Engineered Mouse Model of Lung Cancer by Micro-CT and Automated Analysis1
Barck, Kai H.; Bou-Reslan, Hani; Rastogi, Ujjawal; Sakhuja, Timothy; Long, Jason E.; Molina, Rafael; Lima, Anthony; Hamilton, Patricia; Junttila, Melissa R.; Johnson, Leisa; Carano, Richard A.D.
Genetically engineered mouse models (GEMMs) of lung cancer closely recapitulate the human disease but suffer from the difficulty of evaluating tumor growth by conventional methods. Herein, a novel automated image analysis method for estimating the lung tumor burden from in vivo micro-computed tomography (micro-CT) data is described. The proposed tumor burden metric is the segmented soft tissue volume contained within a chest space region of interest, excluding an estimate of the heart volume. The method was validated by comparison with previously published manual analysis methods and applied in two therapeutic studies in a mutant K-ras GEMM of non¢small cell lung carcinoma. Mice were imaged by micro-CT pre-treatment and stratified into four treatment groups: an antibody inhibiting vascular endothelial growth factor (anti-VEGF), chemotherapy, combination of anti-VEGF and chemotherapy, or control antibody. In the first study, post-treatment imaging was performed 4 weeks later. In the second study, mice were scanned serially on a high-throughput scanner every 2 weeks for 8 weeks during treatment. In both studies, the automated tumor burden estimates were well correlated with manual metrics (r value range: 0.83-0.93, P < .0001) and showed a similar, significant reduction in tumor growth in mice treated with anti-VEGF alone or in combination with chemotherapy. Given the fully automated nature of this technique, the proposed analysis method can provide a valuable tool in preclinical drug research for screening and randomizing animals into treatment groups and evaluating treatment efficacy in mouse models of lung cancer in a highly robust and efficient manner. PMID:25926079
A Twenty-First Century View of Evolution: Genome System Architecture, Repetitive DNA, and Natural Genetic Engineering
NASA Astrophysics Data System (ADS)
Shapiro, James A.
It is essential for nonbiologists to understand that evolutionary theory based on random mutation of autonomous genes is far from the last word on how genomes have changed in the course of biological evolution. The last 50 years of molecular genetics have produced an abundance of new discoveries and data that make it useful to revisit some basic concepts and assumptions in our thinking about genomes and evolution. Chief among these observations are the complex modularity of genome organization, the biological ubiquity of mobile and repetitive DNA sequences, and the fundamental importance of DNA rearrangements in the evolution of sequenced genomes. This review will take a broad overview of these developments and suggest some new ways of thinking about genomes as sophisticated informatic storage systems and about evolution as a systems engineering process.
Reverse genetic engineering of the human rhinovirus serotype 16 genome to introduce an antibody-detectable tag.
Walker, Erin J; Jensen, Lora M; Ghildyal, Reena
The ability to accurately detect viral proteins during infection is essential for virology research, and the lack of specific antibodies can make this detection difficult. Reverse genetic engineering of virus genomes to alter the wild-type genome is a powerful technique to introduce a detectable tag onto a viral protein. Here we outline a method to incorporate an influenza hemagglutinin epitope tag onto the 2A protease of HRV16. The method uses site-directed mutagenesis PCR to introduce the sequence for the HA antigen onto either the C or N termini of 2A protease while keeping the relevant internal cleavage sites intact. The new viral product is then cloned into a wild-type HRV16 plasmid and transfected into Ohio Hela cells to produce recombinant virus. PMID:25261314
Putting Synthesis into Biology ¢ A Viral View of Genetic Engineering Through de novo Gene and Genome synthesis
Mueller, Steffen; Coleman, J. Robert; Wimmer, Eckard
The rapid improvements in DNA synthesis technology hold the potential to revolutionize biosciences in the near future. Traditional genetic engineering methods are template dependent and make extensive but laborious use of site-directed mutagenesis to explore the impact of small variations on an existing sequence ōthemeö. De novo gene and genome synthesis frees the investigator from the restrictions of the pre-existing template and allows for the rational design of any conceivable new sequence theme. Viruses, being amongst the simplest replicating entities, have been at the forefront of the advancing biosciences since the dawn of molecular biology. Viral genomes, especially those of RNA viruses, are relatively short, often less than 10,000 bases long, making them amenable to whole genome synthesis with the currently available technology. For this reason viruses are once again poised to lead the way in the budding field of synthetic biology ¢ for better or worse. PMID:19318214
Development of wireless batteryless implantable blood pressure-EKG-core body temperature sensing microsystem for genetically engineered mice real time monitoring
Darrin J. Young
Two wireless implantable microsystems; one for blood pressure sensing and one for EKG and core body temperature sensing; are developed for untethered genetically engineered mice real-time monitoring. A flat silicone blood pressure sensing cuff and a low power ASIC are employed to form a novel wireless less-invasive blood pressure monitoring microsystem, which demonstrates a packaged system weight of 130 mg
Cloning and mutagenesis: tinkering with the order of Engineering the replication of target DNA through cloning, or changing its genetic code through mutations,
Cai, Long
Cloning and mutagenesis: tinkering with the order of things Engineering the replication of target DNA through cloning, or changing its genetic code through mutations, are detail-oriented processes whose foibles can spell disaster. Caitlin Smith looks at some new tools and techniques that may smooth
Neural Networks and Genetic Algorithms in Materials Science and Engineering, 2006 January 1113, 2006, Tata McGrawHill Publishing Company Ltd., India
Cambridge, University of
Neural Networks and Genetic Algorithms in Materials Science and Engineering, 2006 January 11, Shibpur, Howrah, India Neural Networks in Materials Science: The Importance of Uncertainty H. K. D. H rela- tionships and structure within vast arrays of illŁunderstood data. The neural network method
[Genetic engineering of microbial metabolic pathway for production of advanced biodiesel].
Fu, Ai-Si; Liu, Ran; Zhu, Jing; Liu, Tian-Gang
Biodiesel is a renewable biofuel and alternative diesel, but the first generation of biodiesel, which has many defects in properties and in production methods, mainly comes from the chemical transesterification of triglyceride from plant oil. With the fast development in the field of synthetic biology and metabolic engineer-ing, the researchers can choose suitable microbes and engineer its metabolic pathways, such as fatty acid bio-synthesis pathway and isoprenoid biosynthesis pathway, to directly produce the second generation of advanced biodiesel---long chain hydrocarbons, which have better properties and quality using the newest biotechnology techniques. In this review, we summarized the research progress about microbial production of advanced bio-diesel and also pointed the deficiencies and future direction in this new field. PMID:21993287
Properties of a Genetically Engineered G Domain of Elongation Factor Tu
Andrea Parmeggiani; Guido W. M. Swart; Kim K. Mortensen; Michael Jensen; Brian F. C. Clark; Luciana Dente; Riccardo Cortese
The G domain of elongation factor Tu (EF-Tu), representing the N-terminal half of the factor according to its three-dimensional model traced at high resolution, has been isolated by genetic manipulation of tufA and purified to homogeneity. The G domain, whose primary structure shares homology with the eukaryotic protein p21, is capable of supporting the basic activities of the intact molecule
Construction and optimization of a family of genetically encoded metabolite sensors by semirational protein engineering
Karen Deuschle; Sakiko Okumoto; Marcus Fehr; Loren L. Looger; Leonid Kozhukh; Wolf B. Frommer
A family of genetically-encoded metabolite sensors has been constructed using bacterial periplasmic binding proteins (PBPs) linearly fused to protein fluorophores. The ligand-induced conformational change in a PBP allosterically regulates the relative distance and orientation of a fluorescence resonance energy transfer (FRET)-compatible protein pair. Ligand binding is transduced into a macroscopic FRET observable, providing a reagent for in vitro and in
Engineering modular and orthogonal genetic logic gates for robust digital-like synthetic biology
Baojun Wang; Nicolas Joly; Richard I Kitney; Martin Buck
Modular and orthogonal genetic logic gates are essential for building robust biologically based digital devices to customize cell signalling in synthetic biology. Here we constructed an orthogonal AND gate in Escherichia coli using a novel hetero-regulation module from Pseudomonas syringae. The device comprises two co-activating genes hrpR and hrpS controlled by separate promoter inputs, and a ?54-dependent hrpL promoter driving
Genetic transformation of Brassica campestris var. rapa protoplasts with an engineered cauliflower mosaic virus genome
Jerzy Paszkowski; Barbara Pisan; Raymond D. Shillito; Thomas Hohn; Barbara Hohn; Ingo Potrykus
A hybrid Cauliflower Mosaic Virus (CaMV) genome containing a selectable marker gene was constructed by replacing the gene VI coding region with the aminoglycoside (neomycin) phosphotransferase type II [APH(3')II] gene from Tn5. This modified viral genome was tested for its infectivity both in planta and in a protoplast transformation system of Brassica campestris var. rapa. Stable, genetically transformed cell lines
Restoration of fertility by antisense RNA in genetically engineered male sterile tobacco plants
Thomas Schm³lling; Horst R÷hrig; Silke Pilz; Richard Walden; Jeff Schell
Transgenic tobacco plants (Nicotiana tabacum L.) expressing the rolC gene of Agrobacterium rhizogenes under the transcriptional control of the 35S RNA promoter are male sterile. When these plants are genetically crossed with others containing the rolC gene linked in antisense orientation to the 35S RNA promoter, hybrid progeny display restoration of male fertility. Moreover, hybrid progeny are revertant for other
Genetically engineered vegetables expressing proteins from Bacillus thuringiensis for insect resistance: successes, disappointments, challenges and ways to move forward.
Shelton, Anthony M
Genetically engineered (GE) insect-resistant crops that express proteins from Bacillus thuringiensis (Bt) have been widely adopted in the two field crops currently commercially available, Bt cotton and Bt corn. However, the development and commercialization of Bt vegetables has lagged in comparison, which is unfortunate since vegetables tend to be insecticide-intensive crops due to high pest pressure and cosmetic standards required for the market. While it is often stated that consumer choice has played a major role in companies avoiding development of Bt vegetables, this concept requires re-evaluation. In market studies in North America when consumers have been provided basic information about Bt genetic engineering, then given a choice between Bt and conventional sweet corn, they have often preferred the former. Likewise, 77% of consumers in a US survey said they would likely purchase foods produced through biotechnology for their ability to reduce pesticide use. Presently, however, the only commercialized Bt vegetable is sweet corn. Perhaps more critical obstacles to Bt vegetables are their relatively smaller acreages and the cost of government biosafety regulations that inadvertently favor large acreage of field crops because companies can obtain a better return on investment. In developing countries, private-public partnerships may provide the vehicle to bring Bt vegetables to market. However, these can be subverted by misinformation from anti-biotech campaigns, as is the case with Bt eggplant in India. Without the use of Bt vegetables as a tool for integrated pest management, farmers and the general public will not be able to realize the substantial environmental and economic benefits that have been well documented with Bt cotton and Bt corn. PMID:22538234
Comparison of immune responses to different foot-and-mouth disease genetically engineered vaccines in guinea pigs.
Yao, Qingxia; Qian, Ping; Huang, Qinfeng; Cao, Yi; Chen, Huanchun
The P12A3C gene from FMDV (serotype O) encoding the capsid precursor protein, and the highly immunogenic gene FHG, which encodes multiple epitopes of FMDV capsid proteins, were inserted into eukaryotic expression vectors to compare different candidate genetically engineered vaccines for foot-and-mouth disease (FMD). A modified live pseudorabies virus (MLPRV) was also used to deliver P12A3C. Guinea pigs were inoculated intramuscularly with the candidate vaccines to compare the ability to elicit immunity of the DNA vector and a live viral vector. An indirect enzyme-linked immunosorbent assay (iELISA), virus-neutralization test and lymphoproliferation assay were used to detect antibody and cellular responses. The group immunized with P12A3C delivered by MLPRV produced significantly greater antibody and cellular responses indicating that MLPRV has a greater ability to mediate exogenous gene delivery than the plasmid DNA vector. Comparison of the immune responses induced by P12A3C and FHG, which were both mediated by DNA plasmids, showed that FHG and P12A3C elicited similar cellular responses, while P12A3C induced higher antibody levels, suggesting that P12A3C is a more powerful immunogen than FHG. In challenge experiments, guinea pigs vaccinated with P12A3C delivered by MLPRV were protected fully from FMDV challenge, whereas guinea pigs vaccinated with P12A3C or FHG delivered by DNA plasmid were only protected partially. This study provides a basis for future construction of a genetically engineered vaccine for FMDV. PMID:17963851
A Science, Engineering and Technology (SET) Approach Improves Science Process Skills in 4-H Animal Science Participants
ERIC Educational Resources Information Center
Clarke, Katie C.
A new Science, Engineering and Technology (SET) approach was designed for youth who participated in the Minnesota State Fair Livestock interview process. The project and evaluation were designed to determine if the new SET approach increased content knowledge and science process skills in participants. Results revealed that youth participants notģ
Guidelines for the Genotyping of Mice and Rats The specific genetic identification of genetically engineered animals in a litter is critical to the
Bandettini, Peter A.
, fecal or oral samples1-12 . Larger amounts of DNA are required for Southern Blot determination for the procedure14 . DNA prepared from tail biopsies is suitable for analysis by either Southern Blot or PCR