The present inventors successfully produced rice dwarf mutant d61 and also isolated the OsBRI1 gene which corresponds to a region in the d61 locus. OsBRI1 is found to increase plant brassinosteroid sensitivity. Moreover, the present inventors showed that OsBRI1 functions in growth and development process of rice, such as, internode elongation by inducing internode cell elongation and the inclination of the lamina joint. By introducing antisense nucleotides or dominant negative of OsBRI1, the present inventors produced transgenic rice plants whose phenotype was modified.

Research in plant molecular biology has advanced dramatically in recent years and is necessary for the analysis of various physiological phenomena. Dwarfism caused by artificial modification of grass type, especially the control of elongation growth, prevents plants from lodging due to overgrowth caused by over fertilization. This prevention of lodging was demonstrated in Mexican wheat during the "Green Revolution" and in miracle rice (IR-8) developed by the International Rice Research Center. Furthermore, in the case of cultivation at high density, such as rice cultivation, yields are expected to increase as a result of the increase in the amount of sun light each plant receives due to the formation of upright leaves. Moreover, these modifications are very important breeding targets because they may result in yield increases and also increase the efficiency of plant growth maintenance. However, current breeding methods cannot artificially modify plant morphology.
Dwarfism is an abnormal growth caused by mutation in genes involved in controlling normal elongation growth. Plant elongation growth is the result of accumulation of cell division and cell elongation. Cell division and cell elongation are controlled by complex effects caused by various factors, such as, exogenous environmental factors including temperature and light and endogenous environmental factors including plant hormones. Therefore, it is predicted that many genes, such as those related to plant hormone biosynthesis and hormone receptors directly and those related to the control of the expression of these genes, are involved in the dwarfism (Sakamoto et al. (2000) Kagaku to Seibutsu, 38: 131–139).
Almost all modern cultivars of japonica rice develop 15–16 phytomers, consisting of leaves, axillary buds, and short or elongated internodes, during the vegetative stage. After the shoot meristem shifts from the vegetative to the reproductive phase, the reproductive meristem develops about 10 phytomers consisting of undeveloped leaf, an elongated internode, and an axillary which develops into the primary rachis branch. The phytomers formed in the vegetative stage can be classified into three types in terms of the morphology of the internode (Suetsugu, Isao. (1968) Japan. J. Crop Sci. 37, 489–498). The first type is developed in the juvenile phase and form undifferentiated nodes and internodes. After the shoot apical meristem (SAM) shifts from the juvenile to the adult phase, the nodal plate of the second type differentiates and the central part of the internode thereof decays to produce an air space. The third type contains long elongated internodes as a result of growth from the intercalary meristem.
Phytomers of type 1 are produced first during vegetative development, followed by type 2 and then type 3 phytomers. Under normal growth conditions, the number of phytomers of each type in many japonica cultivars is 4–5, 6–7, and 4–5. The transition from type 1 to type 2 is strictly regulated. After the serial development of 4–5 type 1 phytomers, depending on the cultivar, the SAM develops type 2 phytomers. However, the transition from type 2 to type 3 does not depend on the number of development of type 2 phytomers. The SAM develops 15–16 phytomers and shifts from the vegetative to the reproductive stage, the type 3 phytomers then start to develop, and the uppermost four or five internodes thereof start to elongate. If the timing of the transition is changed by unusual growth conditions, the number of the type 2 phytomers always affected thereby, but the number of the type 3 phytomer with elongated internode is unchanged (Suetsugu, isao. (1968) Japan. J. Crop Sci: 37, 489–498). This indicates that the transition of the SAM from the vegetative to the reproductive phase in rice induces internode elongation, as well as in Arabidopsis.
However, there is an important difference between rice and Arabidopsis. The elongated internodes in rice are derived from the vegetative SAM while those in Arabidopsis come from the reproductive SAM. In rice, the uppermost four or five internodes develop from the vegetative SAM and initially are indistinguishable from the lower type 2 internodes. When the SAM shifts to the reproductive phase, differentiation into type 3 internodes occurs due to the development of intercalary meristems in the internodes. This synchronicity between the phase change of the SAM and the development of the intercalary meristem leads to the possibility that these processes might be linked by a signal coming from the SAM to the uppermost four or five phytomers when its phase change occurs.
A large number of dwarf mutants of rice have been collected and characterized because of their agronomic importance. These dwarf mutants are categorized into six groups based on the elongation pattern of the upper four to five internodes (FIG. 1; redrawn from Takeda, K. (1974) Bull. Fac. Agr. Hirosaki Univ. 22, 19–30. In rice, each internode is numbered from top to bottom such that the uppermost internode just below the panicle is first). The present inventors can see that in the dn-type mutants the length of each internode is almost uniformly reduced, resulting in an elongation pattern similar to that of the wild type plant. In contrast, the dm-type mutants show specific reduction of the second internode. Similar shortening of a specific internode is also observed in the sh- and d6-type mutants, in which only the uppermost first internode or internodes below the uppermost are shortened, respectively. As these mutants with specifically shortened internodes, such as the dm-, d6-, and sh-types, might be defective in the perception of signals coming from the SAM, they should be especially useful for the study of the mechanism of internode elongation and its relationship to changes in the SAM.
Brassinosteroids (BRs) are plant growth-promoting natural products that are required for plant growth and development. There are only a few reports on the physiological effects of brassinosteroids in the growth and development of rice and other plants of the Gramineae family. Physiological researches indicate that exogenous brassinosteroids alone, or in combination with auxin, enhance bending of the lamina joint in rice. The lamina joint has been used for a sensitive bioassay of brassinosteroids (Maeda, E. (1965) Physiol. Plant. 18, 813–827; Wada, K. et al. (1981) Plant and Cell Physiol. 22, 323–325; Takeno, K. and Pharis, R. P. (1982) Plant Cell Physiol. 23, 1275–1281), because of high sensitivity thereof to brassinosteroids. In etiolated wheat seedlings treatment with brassinolide or its derivative, castasterone, stimulates unrolling of the leaf blades (Wada, K. et al. (1985) Agric. Biol. Chem. 49, 2249–2251). Treatment with low or high concentrations of brassinosteroids promotes or inhibits the growth of roots in rice, respectively (Radi, S. H. and Maeda, E. (1988) J. Crop Sci. 57, 191–198). Brassinosteroids also promote the germination of rice seeds (Yamaguchi, T. et al. (1987) Stimulation of germination in aged rice seeds by pre-treatment with brassinolide. In: Proceeding of the fourteenth annual plant growth regulator society of America Meeting Honolulu. (Cooke A R), pp. 26–27).
Although these results indicate only effects due to exogenous brassinosteroids, not due to endogenous brassinosteroids, they do suggest that endogenous brassinosteroids have an important role in growth and developmental processes in plants of the Gramineae family.
On the other hand, there is some apparent disagreement in the literature as to whether brassinosteroids induce cell elongation in plants of the Gramineae family. That is, brassinolide treatment does not induce elongation of the leaf sheath of rice (Yokota, T. and Takahashi, N. (1986) Chemistry, physiology and agricultural application of brassinolide and related steroids. In: Plant growth substances 1985. (Bopp M, Springer-Verlag, Berlin/Heidelberg/New York) pp.129–138), but it does induce elongation of the coleoptile and mesocotyl in maize (He, R. —Y. et al. (1991) Effects of brassinolide on growth and chilling resistance of maize seedlings. In: Brassinosteroids-Chemistry, Bioactivity and Applications ACS symposium series 474. (Cutler H G C, Yokota T, Adam G, American Chemical Society, Washington D.C.), pp. 220–230).
As shown by brassinosteroids synthesis mutants or brassinosteroids insensitive mutants that show severe dwarfism with abnormal development of organs, the function of brassinolide is known in dicotyledonous plants.
However, little is known about the function of endogenous brassinosteroids in monocotyledonous plants, such as rice or other plants of the Gramineae family.
3. Disclosure of the Invention
The object of the present invention is to provide novel genes involved in brassinosteroid sensitivity from plants, preferably from monocotyledonous plants. Another object of the present invention is to modify plant brassinosteroid sensitivity by controlling the expression of the gene. The modification in plant brassinosteroid sensitivity causes a change in plant morphology. The preferable embodiment of the present invention provides plants with erect leaves which become dwarfed due to the suppression of internode elongation caused by decreased brassinosteroid sensitivity.
By treatment with mutagenesis agent, the present inventors isolated a novel rice dwarf mutant strain d61 (d61-1 and d61-2) which showed lower brassinosteroid sensitivity and had shorter internodes than wild type plants.
Linkage analysis indicated that the d61 locus was highly linked to a gene region that was homologous to Arabidopsis BRI1. The present inventors isolated the gene (OsBRI1), which was homologous to Arabidopsis BRI1 gene, by screening of a rice genomic DNA library. Nucleotide sequence analysis of the OsBRI1 gene from d61-1 and d61-2 mutants indicated that there were single nucleotide substitutions causing amino acid substitutions at different sites in each d61 allele.
Moreover, in order to confirm that the OsBRI1 gene corresponds to the d61 locus, the OsBRI1 gene was introduced into d61 mutants. As a result, the OsBRI1 gene complimented the d61 phenotype and caused the mutant strain to have a wild-type phenotype. Therefore, it was indicated that d61 mutants are caused by loss of function of the OsBRI1 gene. Phenotypic analysis of plants revealed that the OsBRI1 gene functions in various growth and development processes of rice including internode elongation caused by formation of intercalary meristem and induction of internode cell longitudinal elongation, inclination of the lamina joint, and skotomorphogenesis in the dark.
Moreover, in the case where transgenic rice plants with OsBRI1 antisense nucleotide were produced, most transgenic plants produced erect leaves during seedling growth. All of the transgenic plants showed dwarf phenotype of various levels. Plants transformed with OSBRI1 having the dominant negative phenotype showed the same result.
The present invention had been made in view of such findings, and relates to a novel gene involved in plant brassinosteroid sensitivity, the protein encoded by the gene, and production and use of the same. Moreover, the present invention relates to the production of modified plant by controlling expression of the gene.
More specifically, this invention provides:    (1) a DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 2;    (2) the DNA of (1), wherein the DNA is a cDNA or a genomic DNA;    (3) the DNA of (1), wherein the DNA comprises a coding region of the nucleotide sequence of SEQ ID NO: 1 or 3;    (4) a DNA encoding a protein which has 55% or more homology to the amino acid sequence of SEQ ID NO: 2 and which is functionally equivalent to a protein comprising the amino acid sequence of SEQ ID NO: 2, the DNA being selected from the group consisting of            (a) a DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 2 in which one or more amino acids are substituted, deleted, added, and/or inserted; and        (b) a DNA hybridizing under stringent conditions with a DNA comprising the nucleotide sequence of SEQ ID NO: 1 or 3;            (5) the DNA of (4), wherein the DNA encodes a protein having a function selected from the group consisting of a function of increasing brassinosteroid sensitivity in a plant, a function of inducing elongation of internode cells of a stem of a plant, a function of positioning microtubules perpendicular to the direction of elongation in an internode of a stem of a plant, a function of suppressing elongation of an internode of a neck of a plant, and a function of increasing inclination of a lamina of a plant;    (6) the DNA of (4) or (5), wherein the DNA is derived from a monocotyledonous plant;    (7) the DNA of (6), wherein the DNA is derived from a plant of the Gramineae family;    (8) a DNA encoding an antisense RNA complementary to a transcript of the DNA of any one of (1) to (7);    (9) a DNA encoding an RNA having ribozyme activity which specifically cleaves a transcript of the DNA of any one of (1) to (7);    (10) a DNA which encodes an RNA repressing expression of the DNA of any one of (1) to (7) due to co-suppression when expressed in a plant cell and which has 90% or more homology to the DNA of any one of (1) to (7);    (11) a DNA which encodes a protein having a dominant negative phenotype to that of a protein encoded by the DNA of any one of (1) to (7);    (12) a vector which comprises the DNA of any one of (1) to (7);    (13) a transformed cell which comprises the DNA of any one of (1) to (7) or the vector of (12);    (14) a protein encoded by the DNA of any one of (1) to (7);    (15) a method for producing the protein of (14) the method comprising the steps of culturing the transformed cell of (13) and recovering an expressed protein from the transformed cell or a culture supernatant thereof;    (16) a vector comprising the DNA of any one of (8) to (11);    (17) a transformed plant cell comprising the DNA of any one of (1) to (11) or the vector of (12) or (16);    (18) a transformed plant comprising the transformed plant cell of (17);    (19) a transformed plant which is a progeny or a clone of the transformed plant of (18);    (20) a breeding material of the transformed plant of (18) or (19); and    (21) an antibody which binds to the protein of (14).
The present invention provides a DNA encoding the OsBRI1 protein derived from rice. The nucleotide sequence of OsBRI1 cDNA is shown in SEQ ID NO: 1, the amino acid sequence of the protein encoded by the DNA is shown in SEQ ID NO: 2, and the nucleotide sequence of the genomic DNA of OsBRI1 is shown in SEQ ID NO: 3 (the genomic DNA of SEQ ID NO: 3 consists of one exon with no intron).
The gene of the present invention causes a rice dwarf mutant (d61) which has short internodes and reduced brassinosteroid sensitivity compared to the wild type. Therefore, it is possible to modify plant morphology by controlling the expression of the OsBRI1 gene.
The preferable modification in plant morphology in the present invention includes dwarfism of plants by suppressing expression of the DNA of the present invention. Dwarfism of plants has great value in agriculture and horticulture. For example, reduction of height of plants can reduce the tendency of plants to lodge and can thereby increase seed weights. Moreover, it is possible to increase the number of plant individuals which can be planted per unit area by reducing height of plants and by making plant shape per plant more compact. These plant modifications have great value specifically in the production of crops such as rice, corn, wheat, and such. It is also possible to produce ornamental plants with new aesthetic value by dwarfism of height or culm length of plants. It is also possible to produce miniature vegetables or fruits with new commercial value, such as "bite-size", by dwarfism of them. Other than for industrial plants, dwarfism is important for experimental plants because, for example, dwarf plants are not only more easily handled but they also help utilize experimental space more effectively by decreasing cultivation space.
It is possible to consider that brassinosteroid sensitivity can be increased in brassinosteroid low sensitive plants by expressing the DNA of the present invention in the plants. Thereby, the yield of whole plants may be increased by growing taller plants. Thus, this will be especially useful for increasing yield for whole feed crops.
DNA encoding the OsBRI1 protein of the present invention includes genomic DNA, cDNA, and chemically synthesized DNA. A genomic DNA and cDNA can be prepared according to conventional methods known to those skilled in the art. More specifically, a genomic DNA can be prepared, for example, as follows: (1) extract genomic DNA from plant cells or tissues; (2) construct a genomic library (utilizing a vector, such as plasmid, phage, cosmid, BAC, PAC, and such); (3) spread the library; and (4) conduct colony hybridization or plaque hybridization using a probe prepared based on the DNA encoding a protein of the present invention (e.g., SEQ ID NO: 1 or 3). Alternatively, a genomic DNA can be prepared by PCR, using primers specific to a DNA encoding the protein of the present invention (e.g. SEQ ID NO: 1 or 3). On the other hand, cDNA can be prepared, for example, as follows: (1) synthesize cDNAs based on mRNAs extracted from plant cells or tissues; (2) prepare a cDNA library by inserting the synthesized cDNA into vectors, such as λZAP; (3) spread the cDNA library; and (4) conduct colony hybridization or plaque hybridization as described above. Alternatively, cDNA can be also prepared by PCR.
The present invention includes DNAs encoding proteins functionally equivalent to the OsBRI1 protein of SEQ ID NO: 2. Herein, the term "functionally equivalent to the OsBRI1 protein" means that the object protein has equal functions to those of the OsBRI1 protein of SEQ ID NO: 2, such as, for example, a function of increasing brassinosteroid sensitivity in a plant, a function of inducing elongation of an internode of a stem of a plant, a function of positioning microtubules perpendicular to the direction of elongation in internode cells of a stem of a plant, a function of suppressing elongation of an internode of a neck of a plant, and/or a function of increasing inclination of a lamina of a plant. Such DNA is derived preferably from monocotyledonous plants, more preferably from plants of the Gramineae family, and most preferably from rice.
Examples of such DNAs include those encoding mutants, derivatives, alleles, variants, and homologues comprising the amino acid sequence of SEQ ID NO: 2 wherein one or more amino acids are substituted, deleted, added, and/or inserted.
Examples of methods for preparing a DNA encoding a protein comprising altered amino acids well known to those skilled in the art include the site-directed mutagenesis (Kramer, W. and Fritz, H. -J. (1987) "Oligonucleotide-directed construction of mutagenesis via gapped duplex DNA." Methods in Enzymology, 154: 350–367). The amino acid sequence of a protein may also be mutated in nature due to the mutation of a nucleotide sequence. A DNA encoding proteins having the amino acid sequence of a natural OsBRI1 protein (SEQ ID NO: 2) wherein one or more amino acids are substituted, deleted, and/or added are also included in the DNA of the present invention, so long as they encode a protein functionally equivalent to the natural OsBRI1 protein. Additionally, nucleotide sequence mutants that do not give rise to amino acid sequence changes in the protein (degeneracy mutants) are also included in the DNA of the present invention. The number of nucleotide mutations of the DNA of interest corresponds to, at amino acid level, typically 100 residues or less, preferably 50 residues or less, more preferably 20 residues or less, and still more preferably 10 residues or less (for example, 5 residues or less, or 3 residues or less).
Whether a certain DNA actually encodes a protein which has a function of increasing inclination of a lamina of a plant can be evaluated, for example, by performing a "lamina joint test" for plants in which the expression of the DNA has been suppressed and by comparing the results with those for wild-type plants (See Example 4). The result of the test may also be an index for evaluating brassinosteroid sensitivity in a plant. In order to evaluate whether the DNA encodes a protein which has a function of inducing elongation of an internode of a stem of a plant, a function of positioning microtubules perpendicular to the direction of elongation in internode cells of a stem of a plant, or a function of suppressing elongation of an internode of a neck of a plant, the morphology of the internode cell of the plant in which expression of the DNA has been suppressed can be observed to be compared with that of wild type (See Examples 2 and 3).
A DNA encoding a protein functionally equivalent to the OsBRI1 protein described in SEQ ID NO: 2 can be produced, for example, by methods well known to those skilled in the art including: methods using hybridization techniques (Southern, E. M. (1975) Journal of Molecular Biology, 98, 503); and polymerase chain reaction (PCR) techniques (Saiki, R. K. et al. (1985) Science, 230, 1350–1354; Saiki, R. K. et al. (1988) Science, 239, 487–491). That is, it is routine for a person skilled in the art to isolate a DNA with high homology to the OsBRI1 gene from rice and other plants using the OsBRI1 gene (SEQ ID NO: 1 or 3) or parts thereof as a probe, and oligonucleotides hybridizing specifically to the gene as a primer. Such DNA encoding proteins functionally equivalent to the OsBRI1 protein, obtainable by hybridization techniques or PCR techniques, are included in the DNA of this invention.
Hybridization reactions to isolate such DNAs are preferably conducted under stringent conditions. Stringent hybridization conditions of the present invention include conditions such as: 6 M urea, 0.4% SDS, and 0.5×SSC; and those which yield a similar stringency with the conditions. DNAs with higher homology are expected to be isolated efficiently when hybridization is performed under conditions with higher stringency, for example, 6 M urea, 0.4% SDS, and 0.1×SSC. Those DNAs isolated under such conditions are expected to encode a protein having a high amino acid level homology with OsBRI1 protein (SEQ ID NO: 2). Herein, "high homology" means an identity of at least 55% or more, more preferably 70% or more, and most preferably 90% or more (e.g., 95% or more), between full-length of amino acids.
The degree of homology of one amino acid sequence or nucleotide sequence to another can be determined by following the algorithm BLAST by Karlin and Altschul (Proc. Natl. Acad. Sci. USA, 90: 5873–5877, 1993). Programs such as BLASTN and BLASTX were developed based on this algorithm (Altschul et al. J. Mol. Biol. 215: 403–410, 1990). To analyze a nucleotide sequences according to BLASTN based on BLAST, the parameters are set, for example, as score=100 and word length=12. On the other hand, parameters used for the analysis of amino acid sequences by the BLASTX based on BLAST include, for example, score=50 and word length=3. Default parameters of each program are used when using BLAST and Gapped BLAST program. Specific techniques for such analysis are known in the art.
The DNA of the present invention can be used, for example, to prepare recombinant proteins, produce transformed plants with phenotypes altered by controlling expression thereof as described above, and so on.
A recombinant protein is usually prepared by inserting a DNA encoding a protein of the present invention into an appropriate expression vector, introducing said vector into an appropriate cell, culturing the transformed cells, and purifying expressed proteins.
A recombinant protein can be expressed as a fusion protein with other proteins so as to be easily purified, for example, as a fusion protein with maltose binding protein in Escherichia coli (New England Biolabs, USA, vector pMAL series), as a fusion protein with glutathione-S-transferase (GST) (Amersham Pharmacia Biotech, vector pGEX series), or tagged with histidine (Novagen, pET series). The host cell is not limited so long as the cell is suitable for expressing the recombinant protein. It is possible to utilize yeasts or various animal, plant, or insect cells besides the above described E. coli. A vector can be introduced into a host cell by a variety of methods known to one skilled in the art. For example, a transformation method using calcium ions (Mandel, M. and Higa, A. (1970) Journal of Molecular Biology, 53, 158–162; Hanahan, D. (1983) Journal of Molecular Biology, 166, 557–580) can be used to introduce a vector into E. coli. A recombinant protein expressed in host cells can be purified and recovered from the host cells or the culture supernatant thereof by known methods. When a recombinant protein is expressed as a fusion protein with maltose binding protein or other partners, the recombinant protein can be easily purified by affinity chromatography.
The resulting protein can be used to prepare an antibody that binds to the protein. For example, a polyclonal antibody can be prepared by immunizing immune animals, such as rabbits, with a purified protein of the present invention or its portion, collecting blood after a certain period, and removing clots. A monoclonal antibody can be prepared by fusing myeloma cells with the antibody-forming cells of animals immunized with the above protein or its portion, isolating a monoclonal cell expressing a desired antibody (hybridoma), and recovering the antibody from the cell. The obtained antibody can be utilized to purify or detect a protein of the present invention. Accordingly, the present invention includes antibodies that bind to proteins of the invention.
In order to produce a transformed plant in which DNAs of the present invention are expressed, a DNA encoding a protein of the present invention is inserted into an appropriate vector; the vector is then introduced into a plant cell; and finally, the resulting transformed plant cell is regenerated.
On the other hand, a transformed plant with suppressed expression of DNAs of the present invention can be created using DNA that represses the expression of a DNA encoding a protein of the present invention: wherein the DNA is inserted into an appropriate vector, the vector is introduced into a plant cell, and then, the resulting transformed plant cell is regenerated. The phrase "suppression of expression of DNA encoding a protein of the present invention" includes suppression of gene transcription as well as suppression of translation into protein. It also includes not only the complete inability of expression of DNA but also reduction of expression.
The expression of a specific endogenous gene in plants can be repressed by methods utilizing antisense technology, the methods which are commonly used in the art. Ecker et al. were the first to demonstrate the antisense effect of an antisense RNA introduced by electroporation in plant cells by using the transient gene expression method (J. R. Ecker and R. W. Davis (1986) Proc. Natl. Acad. Sci. USA 83: 5372). Thereafter, the target gene expression was reportedly reduced in tobacco and petunias by expressing antisense RNAs (A. R. van der Krol et al. (1988) Nature 333: 866). The antisense technique has now been established as a means to repress target gene expression in plants.
Multiple factors are required for antisense nucleic acid to repress the target gene expression. These include, inhibition of transcription initiation by triple strand formation; suppression of transcription by hybrid formation at the site where the RNA polymerase has formed a local open loop structure; transcription inhibition by hybrid formation with the RNA being synthesized; suppression of splicing by hybrid formation at the junction between an intron and an exon; suppression of splicing by hybrid formation at the site of spliceosome formation; suppression of mRNA translocation from the nucleus to the cytoplasm by hybrid formation with mRNA; suppression of splicing by hybrid formation at the capping site or at the poly A addition site; suppression of translation initiation by hybrid formation at the binding site for the translation initiation factors; suppression of translation by hybrid formation at the site for ribosome binding near the initiation codon; inhibition of peptide chain elongation by hybrid formation in the translated region or at the polysome binding sites of mRNA; and suppression of gene expression by hybrid formation at the sites of interaction between nucleic acids and proteins. These factors repress the target gene expression by inhibiting the process of transcription, splicing, or translation (Hirashima and Inoue, "Shin Seikagaku Jikken Koza (New Biochemistry Experimentation Lectures) 2, Kakusan (Nucleic Acids) IV, Idenshi No Fukusei To Hatsugen (Replication and Expression of Genes)", Nihon Seikagakukai Hen (The Japanese Biochemical Society), Tokyo Kagaku Dozin, pp. 319–347, (1993)).
An antisense sequence of the present invention can repress the target gene expression by any of the above mechanisms. In one embodiment, if an antisense sequence is designed to be complementary to the untranslated region near the 5′ end of the gene's mRNA, it will effectively inhibit translation of a gene. It is also possible to use sequences complementary to the coding regions or to the untranslated region on the 3′ side. Thus, the antisense DNA used in the present invention includes DNA having antisense sequences against both the untranslated regions and the translated regions of the gene. The antisense DNA to be used is connected downstream from an appropriate promoter, and, preferably, a sequence containing the transcription termination signal is connected on the 3′ side. The DNA thus prepared can be transfected into the desired plant by known methods. The sequence of the antisense DNA is preferably a sequence complementary to the endogenous gene of the plant to be transformed or a part thereof, but it need not be perfectly complementary so long as it can effectively inhibit the gene expression. The transcribed RNA is preferably at least 90%, and most preferably at least 95% complementary to the transcribed products of the target gene. Sequence complementarity may be determined using the above-described search.
In order to effectively inhibit the expression of the target gene by means of an antisense sequence, the antisense DNA should be at least 15 nucleotides long, preferably at least 100 nucleotides long, and more preferably at least 500 nucleotides long. The antisense DNA to be used is generally shorter than 5 kb, and preferably shorter than 2.5 kb.
DNA encoding ribozymes can also be used to repress the expression of endogenous genes. A ribozyme is an RNA molecule that has catalytic activity. There are many ribozymes having various activities. Research on ribozymes as RNA cleaving enzymes has enabled the design of a ribozyme that site-specifically cleaves RNA. While some ribozymes of the group I intron type or the mRNA contained in RNaseP consist of 400 nucleotides or more, others belonging to the hammerhead type or the hairpin type have an activity domain of about 40 nucleotides (Makoto Koizumi and Eiko Ohtsuka, (1990) Tanpakushitsu Kakusan Kohso (Nucleic acid, Protein, and Enzyme), 35: 2191).
The self-cleavage domain of a hammerhead type ribozyme cleaves at the 3′ side of C15 of the sequence G13U14C15. Formation of a nucleotide pair between U14 and A at the ninth position is considered important for the ribozyme activity. Furthermore, it has been shown that the cleavage also occurs when the nucleotide at the 15th position is A or U instead of C (M.. Koizumi et al., (1988) FEBS Lett. 228: 225). If the substrate binding site of the ribozyme is designed to be complementary to the RNA sequences adjacent to the target site, one can create a restriction-enzyme-like RNA cleaving ribozyme which recognizes the sequence UC, UU, or UA within the target RNA (M. Koizumi et al., (1988) FEBS Lett. 239: 285; Makoto Koizumi and Eiko Ohtsuka, (1990) Tanpakushitsu Kakusan Kohso (Protein, Nucleic acid, and Enzyme), 35: 2191; M. Koizumi et al., (1989) Nucleic Acids Res. 17: 7059). For example, in the coding region of the OsBRI1 gene (SEQ ID NO: 1 or 3), there is a plurality of sites that can be used as the ribozyme target.
The hairpin type ribozyme is also useful in the present invention. A hairpin type ribozyme can be found, for example, in the minus strand of the satellite RNA of tobacco ringspot virus (J. M. Buzayan, Nature 323: 349 (1986)). This ribozyme has also been shown to target-specifically cleave RNA (Y. Kikuchi and N. Sasaki, (1992) Nucleic Acids Res. 19: 6751; Yo Kikuchi, (1992) Kagaku To Seibutsu (Chemistry and Biology) 30: 112).
The ribozyme designed to cleave the target is fused with a promoter, such as the cauliflower mosaic virus 35S promoter, and with a transcription termination sequence, so that it will be transcribed in plant cells. However, if extra sequences have been added to the 5′ end or the 3′ end of the transcribed RNA, the ribozyme activity can be lost. In this case, one can place an additional trimming ribozyme, which functions in cis to perform the trimming on the 5′ or the 3′ side of the ribozyme portion, in order to precisely cut the ribozyme portion from the transcribed RNA containing the ribozyme (K. Taira et al. (1990) Protein Eng. 3: 733; A. M. Dzaianott and J. J. Bujarski (1989) Proc. Natl. Acad. Sci. USA 86: 4823; C. A. Grosshands and R. T. Cech (1991) Nucleic Acids Res. 19: 3875; K. Taira et al. (1991) Nucleic Acid Res. 19: 5125). Multiple sites within the target gene can be cleaved by arranging these structural units in tandem to achieve greater effects (N. Yuyama et al. (1992) Biochem. Biophys. Res. Commun. 186: 1271). By using such ribozymes, it is possible to specifically cleave the transcripts of the target gene in the present invention, thereby repressing the expression of said gene.
Endogenous gene expression can also be repressed by co-suppression through the transformation by DNA having a sequence identical or similar to the target gene sequence. "Co-suppression" refers to the phenomenon in which, when a gene having a sequence identical or similar to the target endogenous gene sequence is introduced into plants by transformation, expression of both the introduced exogenous gene and the target endogenous gene becomes repressed. Although the detailed mechanism of co-suppression is unknown, it is frequently observed in plants (Curr. Biol. (1996) 7: R793 (1997), Curr. Biol. 6: 810). For example, if one wishes to obtain a plant body in which the OsBRI1 gene is co-repressed, the plant in question can be transformed with a vector DNA designed so as to express the OsBRI1 gene or DNA having a similar sequence to select a plant having the OsBRI1 mutant character, e.g., a plant with suppressed internode elongation, among the resultant plants. The gene to be used for co-suppression does not need to be completely identical to the target gene, but it should have at least 70% or more sequence identity, preferably 80% or more sequence identity, and more preferably 90% or more (e.g., 95% or more) sequence identity. Sequence identity may be determined by above-described search.
In addition, endogenous gene expression in the present invention can also be repressed by transforming the plant with a gene having the dominant negative phenotype of the target gene. Herein, "a DNA encoding the protein having the dominant negative phenotype" refers to a DNA encoding a protein which, when the DNA is expressed, can eliminate or reduce the activity of the protein encoded by the endogenous gene of the present invention inherent to the plant. Preferably, it is a DNA encoding the peptide (e.g., peptide which contains from 739 to 1035 residues of amino acids of SEQ ID NO: 2 or peptides of another protein equivalent to the peptide) which lacks the N-terminal region but contains the kinase region of the protein of the present invention. Whether the DNA of interest has the function to eliminate or enhance activity of the endogenous gene of the present invention can be determined, as mentioned above, by whether the DNA of interest eliminates or reduces a function of increasing brassinosteroid sensitivity in a plant, a function of inducing elongation of an internode of a stem of a plant, a function of positioning microtubules perpendicular to the direction of elongation in internode cells of a stem of a plant, a function of suppressing elongation of an internode of a neck of a plant, and/or a function of increasing inclination of a lamina of a plant.
Vectors used for the transformation of plant cells are not limited as long as the vector can express inserted genes in plant cells. For example, vectors comprising promoters for constitutive gene expression in plant cells (e.g., califlower mosaic virus 35S promoter); and promoters inducible by exogenous stimuli can be used. The term "plant cell" used herein includes various forms of plant cells, such as cultured cell suspensions, protoplasts, leaf sections, and callus.
A vector can be introduced into plant cells by known methods, such as the polyethylene glycol method, electroporation, Agrobacterium mediated transfer, and particle bombardment. Plants can be regenerated from transformed plant cells by known methods depending on the type of the plant cell (Toki et al., (1995) Plant Physiol. 100:1503–1507). For example, transformation and regeneration methods for rice plants include: (1) introducing genes into protoplasts using polyethylene glycol, and regenerating the plant body (suitable for indica rice cultivars) (Datta, S. K. (1995) in "Gene Transfer To Plants", Potrykus I and Spangenberg Eds., pp66–74); (2) introducing genes into protoplasts using electric pulse, and regenerating the plant body (suitable for japonica rice cultivars) (Toki et al (1992) Plant Physiol. 100, 1503–1507); (3) introducing genes directly into cells by the particle bombardment, and regenerating the plant body (Christou et al. (1991) Bio/Technology, 9: 957–962); (4) introducing genes using Agrobacterium, and regenerating the plant body; and so on. These methods are already established in the art and are widely used in the technical field of the present invention. Such methods can be suitably used for the present invention.
Once a transformed plant, wherein the DNA of the present invention is introduced into the genome, is obtained, it is possible to gain descendants from that plant body by sexual or vegetative propagation. Alternatively, plants can be mass-produced from breeding materials (for example, seeds, fruits, ears, tubers, tubercles, tubs, callus, protoplast, etc.) obtained from the plant, as well as descendants or clones thereof. Plant cells transformed with the DNA of the present invention, plant bodies including these cells, descendants and clones of the plant, as well as breeding materials obtained from the plant, its descendant and clones, are all included in the present invention.
The plant of the present invention is preferably a monocotyledonous plant, more preferably a plant of the Gramineae family, and most preferably a rice. The phenotype of the plant of the present invention is different from the wild type phenotype. The phenotypes changed in the plants developed by the present invention include brassinosteroid sensitivity of a plant, plant growth such as internode cell elongation of the plant stem and internode elongation of the ear, inclination of leaves, and the positioning of microtubules perpendicular to the direction of internode cell elongation in the plant stem.

Gene concerning brassinosteroid-sensitivity of plants and utilization thereof

National Institute of Agrobiological Sciences


A system includes a mode module and an energy module. The mode module generates a mode signal based on a temperature of an engine and at least one of a deceleration signal and a regenerative braking signal. The energy module, based on the mode signal, increases cooling of a coolant of the engine during at least one of a deceleration event of a vehicle and a regenerative braking event. The energy module, while increasing the cooling of the coolant, supplies an overvoltage to a cooling pump of the engine.

The background description provided herein is for the purpose of generally presenting the context of the disclosure. Work of the presently named inventors, to the extent it is described in this background section, as well as aspects of the description that may not otherwise qualify as prior art at the time of filing, are neither expressly nor impliedly admitted as prior art against the present disclosure.
A vehicle may include an internal combustion engine (ICE) and an engine control module (ECM). The ECM may control a fuel system and a cooling system of the ICE and one or more regenerative braking systems. The fuel system provides fuel to the ICE and may perform deceleration fuel cut off (DFCO) to conserve fuel. DFCO may include disabling fuel to the ICE. The fuel may be disabled while a driveline (e.g., transmission, driveshaft, axle, and wheels) is engaged to the ICE.
DFCO is used for various reasons. DFCO may be used to provide deceleration (powertrain braking) force when an accelerator of a vehicle is not actuated (e.g., vehicle operator does not press on accelerator pedal). In high elevation (mountainous) areas and/or areas with large variations in elevation, DFCO is used to provide powertrain braking to avoid damage to friction brakes of a vehicle.
DFCO may also be used to prevent damage to a catalytic converter. For example, a throttle position may be calibrated and fixed to provide a minimal amount of air per cylinder (APC) to an engine, thereby providing vehicle deceleration when traveling downhill. Due to the fixed throttle position and/or a manual pull down of a transmission (PRNDL) shifter (e.g., shift into a low gear, such as L1 or L2), the APC levels of the ICE can become too low and cause a misfire. A misfire refers to incomplete combustion of an air/fuel mixture in a cylinder of the engine. This misfire can result in fuel entering and igniting in an exhaust system, which increases the temperature of a catalyst of the catalytic converter. Damage to the catalyst can occur when the temperature of the catalyst exceeds a threshold. By using DFCO, fuel is disabled, which protects the catalyst from misfire events.
DFCO may also be used to increase fuel economy. The efficiency of a gasoline spark ignited engine can be low at minimum combustion (i.e. minimum air and fuel levels) because of pumping losses and other factors. Disabling the fuel is more efficient than reducing the amount of fuel to an ICE.
A cooling system may include an electric cooling pump and an electric cooling fan. The cooling pump circulates coolant through the cooling system including the ICE. The ECM may activate the electric cooling pump by enabling supply of a predetermined voltage (e.g., 12 volts (V)) to the electric cooling pump. The ECM may activate the cooling fan to cool the coolant when the temperature of the coolant is greater than a predetermined temperature.
A regenerative braking system converts kinetic energy generated during engine braking and/or during wheel braking to electrical energy. The electrical energy may be used to charge a vehicle power source and/or used to power electrical components of a vehicle. A regenerative braking system may include one or more generator(s). The generators may apply a braking (i.e., negative) torque on the ICE and/or wheels of a vehicle to perform regenerative braking and produce electrical energy during, for example, deceleration of the ICE and/or vehicle.

Energy control systems and methods for a powertrain of a vehicle


A device for dispensing flexible sheet material, such as paper toweling, from a roll has a feed mechanism including a crank- or lever-operated roller having a drive section and a pressure element engageable with toweling extending around a portion of the roller. The pressure element is spring-biased toward the outer surface of the roller drive section and provides sufficient friction between the roller drive section and the toweling to cause it to be unwound from a roll and advanced for dispensing with out the need for an idler roller which can cause jamming. In one embodiment, a transfer assembly is provided for sequentially dispensing toweling from a primary roll and a reserve roll, after all the toweling has been dispensed from the primary roll. The transfer assembly is mounted for movement between a non-transfer position, where a sensing arm is biased into engagement with the outer surface of toweling on the primary roll to effectively sense the thickness of both the toweling on and the roll core of the primary roll and a pair of transfer legs on a transfer arm are spaced outwardly from the outer surface of the roller, and a transfer position, where the sensing arm engages the outer surface of the primary roll core and the transfer legs push toweling from the reserve roll against the outer surface of the roller and cause it to be fed between the dispensing surface of the pressure element and the roller drive section.

This invention relates to dispensers for rolled sheet material and, in one aspect, to dispensers for paper towel rolls.
Dispensers for rolls of flexible sheet material, such as paper towel rolls, are well known. Paper towel dispensers are widely used in public lavatories to dispense paper toweling for users to dry their hands. Such dispensers typically include either a crank or lever which a user operates to drive a feed mechanism for dispensing the paper toweling. The feed mechanism typically includes a drive roller rotated by a crank or lever and an idler roller, the paper toweling is threaded between these two rollers and the idler roller is spring loaded to provide sufficient friction for rotation of the drive roller to unwind the paper toweling off a roll core. There usually is a relatively small clearance between the drive and idler rollers and the paper toweling can become backed up and cause jamming.
Some paper towel dispensing devices are capable of sequentially dispensing two rolls of paper toweling. This type dispenser is arranged so that, when all the paper toweling from a primary roll has been dispensed, the feed mechanism starts dispensing paper toweling from a reserve roll. Some multiple-roll dispensers have a transfer mechanism including a number of rollers which are arranged to provide tension between the paper toweling being unwound from the primary roll and the feed mechanism and, in response to loss of this tension, effect a transfer of paper toweling from the reserve roll into the feed mechanism. This loss of tension usually occurs when the tail end of the paper toweling has been completely unwound and detached from the primary roll core. However, a loss of tension occasionally can occur during normal use even though there still is paper on the primary roll. For example, if the primary roll core binds and the user is required to use additional force to operate the crank or lever, the primary roll can overrun after the bind is broken loose. This can cause a slack condition which is the same as a loss of tension to the transfer mechanism and it then makes a false transfer, causing paper toweling from the reserve roll to be introduced into the feed mechanism. When this occurs, the feed mechanism can become jammed and no longer capable of dispensing paper toweling or to simultaneously dispense paper toweling from both the primary and reserve rolls.
Some prior constructions which employ a series of rollers for making a transfer from the primary roll to the reserve roll are vulnerable to not effecting a transfer under some conditions which can occur during normal use. Many prior constructions are designed to dispense only one size roll; others are sensitive to the thickness and/or type of the paper toweling being dispensed; others are limited with respect to the size of paper towel roll and/or the roll core size.
Various attempts have been made to minimize a false transfer. For example, U.S. Hedge et al. Pat. No. 4,165,138 discloses a transfer assembly which does not rely on sensing a loss of tension to trigger a transfer. However, the transfer assembly includes two or more rollers with the attendant shortcomings and a number of other parts which increase the overall complexity and cost of construction.


A thermoplastic resin composition, which is flexible and can easily be extruded, is obtained by blending (A) 100 parts by weight of a thermoplastic urethane elastomer with (B) 1-100 parts by weight of a thermoplastic graft copolymer obtained by graft polymerization of vinylidene fluoride with an elastomeric copolymer of at least two principal monomers including at least one fluorine-containing compound, e.g. vinylidene fluoride and chlorotrifluoroethylene, and an unsaturated peroxy compound. Compared with the urethane elastomer (A) itself, this resin composition is better in extrudability, lower in kinetic friction coefficient, weaker in surface tackiness of molded products and higher in resistance to heat aging.

This invention relates to a thermoplastic resin composition which is a blend of a thermoplastic urethane elastomer and a specifically selected thermoplastic fluororesin having flexibility. The resin composition is particularly suitable for extrusion to form, for example, tubes or coverings of electric wires or cables.
Urethane elastomers are widely used as thermoplastic resins having excellent mechanical properties. In particular urethane elastomers having a glass transition temperature lower than room temperature are largely used as extrusion molding materials to form various tubes and coverings of electric wires and cables.
However, from some aspects thermoplastic urethane elastomers have disadvantages too. First, compared with more popular thermoplastic resins such as polyvinyl chloride resins conventional urethane elastomers are generally inferior in extrudability and hence offer greater load to extruders. Therefore, when an urethane elastomer is extruded with an extruder primarily designed for extrusion of other thermoplastic resins it is likely that the extrusion output per unit time and some other items of extrusion conditions are unstable by reason of insufficient power of the extruder.
Thermoplastic urethane elastomers relatively low in hardness have another disadvantage that the extrusion molded products have considerably tacky surfaces. When the products such as tubes or covered wires are left stacked at room temperature the products stick to each other, and in some cases the struck products cannot easily be separated from one another. In industrial practice it is often to apply an antisticking agent in the form of powder or paste to the extruded products of urethane elastomer, but the application of such a powder or paste is troublesome and in many cases raises the need of removing the antisticking agent at the stage of using the extruded products. In some cases the tacky products tend to stick to articles made of different materials and consequently raise certain problems. For example, when a cable having an urethane elastomer covering is used in an industrial robot there is a possibility that the cable sticks to a rack or another cable covered with a different material and consequently breaks as the robot repeats preprogrammed operations.
Besides, as covering materials for electric wires and cables conventional thermoplastic urethane elastomers are not fully satisfactory in resistance to heat aging and in this respect are inferior to conventional fluororesins.

Urethane elastomer based thermoplastic resin composition suitable for extrusion


Shrub rose plant having perfectly rounded, full petaled, fresh tea-scented flowers of a soft creamy yellow that fades to ivory; an attractive, busy growth habit; and high disease resistance.

The present invention relates to a new and distinct variety of rose plant of the shrub class which was originated by me by crossing two unnamed seedlings.
The primary objective of this breeding was to produce a new rose variety having a disease-resistant, strongly-growing plant with blooms of an old-fashioned shape that repeat flowers well. The objective was substantially achieved, along with other desirable improvements, as evidenced by the following unique combination of characteristics that are outstanding in the new variety and that distinguish it from its parents, as well as from all other varieties of which I am aware:
1. Fresh tea-scented fragrance;
2. Attractive, bushy growth;
3. Very disease-resistant; and
4. Perfectly rounded, full-petalled flowers of a soft, creamy yellow that fades to ivory.
Asexual reproduction of this new variety by budding, as performed at David Austin Roses Ltd., Bowling Green Lane, Albrighton, England, shows that the foregoing and all other characteristics and distinctions come true to form and are established and transmitted through succeeding propagations.

Shrub rose plant named xe2x80x98Ausracexe2x80x99


Disclosed is an apparatus and methodology for characterization of small devices. On-chip subtraction of parasitic effects including coupling capacitive effects is provided by way of a rat-race employing a pair of gaps. A device or material being tested is positioned in a test position gap and an output signal is extracted from the rat-race at a position displaced along the rat-race between the test position gap and the other gap to provide subtractive cancellation of any parasitic effects associated with the rat-race and especially associated with the test position gap.

Parasitic effects from measurement fixtures are difficult to handle in microwave characterization of small devices and materials. It is even more challenging when the expected signal level is low. Such devices include magnetroelectronic/spintronic devices and high-impedance devices, for instance a metallic single-walled-carbon-nanotube (mSWNT), a SWNT transistor, a minimum-size deep-submicron metal-oxide-semiconductor (MOS) field-effect-transistor (FET), a sub-micron MOS FET that is operating in sub-threshold region, and a molecular device. Such materials include on-chip biofluids, chemicals and thin films. The microwave characteristics of these devices and materials are of great interest in their development and applications. Considering a metallic SWNT as an example; its high-frequency characteristics are important for potential interconnect and sensor applications. Metallic SWNTs are also considered an ideal, one-dimensional model for fundament condensed matter physics studies. Their high-frequency properties, which correspond to collective Plasmon oscillations, would be a direct verification of Luttinger liquid theory that was proposed to describe one-dimensional material.
A proposed RF transmission line model as illustrated enclosed in the dashed-line box 130 in FIG. 1(c), has often been the focus and foundation for previous mSWNT studies. It has been predicted that mSWNTs have very high characteristic impedance, on the order of 10 kΩ due to high kinetic inductance and includes a large contact resistance component of a few kΩ or higher. Under these conditions, signal attenuation and reflection are very high when currently available microwave measurement systems are used for their characterizations. The difficulty is further complicated by the parasitic effects coming from RF test fixtures, especially the effects from gap coupling capacitor equivalently illustrated at C′p (FIG. 1(c)) and Cp (FIG. 1(d)), in typical measurement arrangements.
The calculated scattering parameters, S21, of the equivalent circuit 140 (FIG. 1(d)) as illustrated in FIGS. 2(a) and 2(b) of several situations exemplify the measurement challenge. At low frequencies, the contact resistance (R) dominates; at high frequencies, the coupling capacitance (Cp) dominates. The inductance effect, which is an indication of transmission line characteristics is indistinguishable from the combined coupling capacitance and contact resistance effects. Moreover, measurement uncertainties, including contact uncertainties from one measurement to another, make it difficult to use various de-embedding or off-chip approaches that have been developed for deep sub-micron CMOS device characterizations. As a result, only limited success has been achieved in the efforts on high-frequency mCNT property and potential application investigations. These efforts include the development of measurement methods for CNT devices. Previously attempted direct scattering parameter measurements, resonant circuit techniques, and heterodyne methods have not been successful in addressing the measurement difficulties for the characterization of the equivalent transmission line model of a mCNT. For instance, the exceptionally large and unique "kinetic" inductance associated with mCNTs has not yet been experimentally verified and characterized.
While various measurement methods for characterizing CNT devices have been developed, no design has emerged that generally encompasses all of the desired characteristics as hereafter presented in accordance with the subject technology.

High-frequency structures for nanoelectronics and molecular electronics


An apparatus includes a dedicated material handling module having a dedicated automated material handling system (AMHS) defines a transport route between a first tool and a second tool selected from a plurality of tools in a fabrication facility. The dedicated AMHS is configured to transport wafer carriers between the first tool and the second tool or vice versa independent of a fabrication facility AMHS that is configured to transport wafer carriers among the plurality of tools.

Modern semiconductor fabrication facilities (fabs) typically use a variety of different tools to perform various fabrication steps on a wafer, such as in the production of integrated circuits (ICs) on silicon wafers. For example, a fab may include tools for performing a variety of different processing and inspection steps such as, e.g., lithography, metrology, etching, ion implantation, deposition, etc. In order to efficiently transport work-in-process (WIP) among the various different tools to perform different fabrication steps on the WIP, a material handling system is generally employed in the fab. Material handling systems in current and next generation fabs typically include an automated material handling system (AMHS), and may also include some amount of manual transport of material by workers in the facility.
Automated material handling systems that are commonly employed in fabrication facilities include, e.g., overhead hoist transports (OHT), rail guided vehicles (RGV), automated guided vehicles (AGV), overhead shuttles (OHS), conveyor systems, and combinations thereof. Regardless of the particular type of AMHS used in the facility, the AMHS will often transport wafers to the various tools in lot containers, called cassettes or carriers, that each hold a plurality of wafers at a time. Commonly employed wafer carriers include mechanical interface (SMIF) pods and front opening unified pods (FOUPs), each of which may hold a plurality of wafers and enable access to individual wafers contained therein by a respective tool's material handler. With the emergence of 300 mm wafers, automated OHT systems which transfer the wafers in FOUPs have become increasingly common.
Each tool's station may generally include one or more material handlers for handling the WIP that is transported to it in lots via the fab AMHS. The material handler at a tool's station may generally include one or more load ports (LP) for loading and unloading FOUPs via the AMHS of the fab, as well as a robotic handling system for removing individual wafers from the FOUP and transferring them to the tool for a particular processing or measurement step.
The material handling system of the fab facility may also have one or more stockers for storing lots, such as, e.g., for temporarily storing wafers due to time deviations among the various tools, or for storage to facilitate transport of the stockers between various fabrication bays. Each stocker may have one or more load ports for loading and unloading FOUPs to and from the AMHS of the fab. The AMHS of a facility may also include a material control system (MCS) for controlling the flow WIP in the facility and issuing commands to the various transport and stocker modules in the AMHS.
By way of example and not by way of limitation, a typical AMHS in a semiconductor fab may utilize an OHT system that includes a track defining a route between various tools and/or stockers in the facility. The track may be located near a ceiling of the facility and have one or more computer-controlled OHT vehicles that travel on the track. Each vehicle may include a gripper for gripping a FOUP from a load port of a stocker or tool and a hoist mechanism that raises and lowers the gripper to raise and lower a FOUP. In a typical operation, the OHT vehicle may be positioned over a load port of a tool or stocker to lower the gripper and retrieve a FOUP from the load port. The hoist mechanism raises the FOUP and the OHT vehicle then transports the FOUP along the track to another load port of a tool or stocker in order to unload the FOUP for storage or a subsequent fabrication step. Each of these lot transfers may be performed according to a transport command received from the fab MCS.
There are currently available several different semiconductor fabrication facility layouts and material transportation architectures for moving wafers between the lithography processing stations and metrology or other inspection stations. Examples of metrology and/or inspection actions frequently performed in such configurations include, but are not limited to, overlay, critical dimension, focus, dose, film thickness, and macro and micro defect inspection. The rapid transportation of WIP from the lithography processing cell to the metrology station is particularly critical to lithographic semiconductor processing because of the possibility of lithography rework, wherein the WIP is returned to a lithography processing cell after metrology or inspection results, e.g., for resist removal and additional lithographic processing if the metrology or inspection results are outside of predetermined limits.
In FIG. 1, one such layout is depicted having a standalone configuration 100, in which a post-lithography metrology station 105a is a separate entity with its own wafer handler 110. A robot 120 within the cell 105b transfers individual wafers between FOUPs at one or more load ports 104 and a processing tool 106. The robot 120 transfers wafers between the cell 105b and the tool 106 via a slit valve 108. The metrology station 105a can be arbitrarily located in the fabrication facility but is often located close to the lithography processing cell 105b. FOUPs can be transferred between the metrology state 105a and cell 105b via the fab's AMHS. This configuration is advantageous in terms of flexibility and cost of ownership as it allows a single metrology station to serve a number of lithography processing cells, relying on the AMHS of the fabrication facility to convey material to and from the station. In this standalone configuration, wafers are generally transferred between the lithography processing cell 105b and the metrology station 105a in cassettes. However, the stand alone configuration 100 suffers from a major drawback of long and varying time to result, while the metrology system 105a waits for the AMHS of the facility to transfer a completed lot from the lithography cell 105b.
In FIG. 2, another such layout is depicted having an integrated metrology configuration 200, in which the metrology station 205a is integrated with a lithography processing cell 205b. As in the configuration 100 shown in FIG. 1, wafers are transferred between FOUPs at one or more load ports 204 and the cell 205b by a robot 220. In this configuration, the robot 220 can directly transfer single wafers between a lithography tool 206 and the metrology station 205a for measurement. A crucial benefit of the integrated configuration 200 is that of time to result since wafers can be individually transferred into the metrology station 205a prior to completion of processing of the whole lot, thereby allowing feedback of results within lot processing time.
In FIG. 3, yet another such layout is depicted having an embedded metrology configuration 300, whereby the metrology station 305a is integrated into the exposure tool 325, and single wafers are transferred from the exposure tool 325 to the metrology station 305a for measurement immediately subsequent to exposure and prior to subsequent processing, such as develop or post exposure bake. This has additional benefits in terms of time to result but suffers from metrology challenges due to low image contrast prior to processing.
Both the integrated configurations and embedded metrology configurations suffer from cost of ownership disadvantages since the metrology station is dedicated to the lithography cell, resulting in substantial idle times for the metrology station during and between lot processing periods. Likewise, the standalone configuration suffers from long and varying time to result because it relies on the facility level AMHS to transfer completed lots form the lithography station.
It is within this context that aspects of the present disclosure arise.

Material handling with dedicated automated material handling system


A basket style article carrier includes a bottom wall, a pair of side walls foldably joined to opposite side edges of the bottom wall, end panels foldably joined to each end of each side wall, a riser panel foldably joined to the inner edge of the end panels at one end of the carrier, a pair of medial panels foldably joined respectively to the inner edges of the end panels at the other end of the carrier, a pair of handle panels foldably joined respectively to the medial panels and secured in flat face contacting relation with the medial panels, cross partition structure foldably joined to each medial panel and a longitudinal reinforcing beam panel foldably joined along its top edge to the bottom edge of one medial panel and secured at one end to one of the medial panels and at the other end to one of the riser panels.

U.S. Pat. No. 3,432,073 issued Mar. 11, 1969 discloses a basket style article carrier which includes a medial strut to which cross partition elements are foldably joined. In this patent, the partition structure is folded downwardly about the upper edge of the medial strut to which the cross partition elements are foldably joined. This patent is devoted to special handle structure and is primarily concerned with affording protection for packaged articles which extends a substantial distance from the bottom parts of the packaged items to the top portions thereof. This patent is not concerned primarily with providing reinforcing medial structure whereby a basket carrier is rendered mechanically strong and substantially rigid from one end to the other particularly along its medial elements.
Also of interest are the following patents:
U.S. Pat. No. 2,687,232 PA1 U.S. Pat. No. 2,820,572 PA1 U.S. Pat. No. 2,905,359 PA1 U.S. Pat. No. 3,105,608 PA1 U.S. Pat. No. 3,850,292 PA1 U.S. Pat. No. 3,917,058 PA1 U.S. Pat. No. 4,311,269 PA1 U.S. Pat. No. 4,313,554 PA1 843,718 British PA1 74 11 248.4 German filed 3/30/75


A vehicular frame assembly includes a mainframe, a support bracket, and a subframe. The support bracket is coupled with the mainframe. The subframe is coupled with the support bracket such that the subframe is stationary with respect to the mainframe. The support bracket is configured for pivotal attachment to a suspension element. Vehicles including a frame assembly having a support bracket are also provided.

Conventional vehicles include a subframe which is attached to a mainframe of the vehicle. The subframe is configured to support body or other components of the vehicle with respect to the mainframe. Some of these conventional vehicles also include wheel suspension components which are also attached to the mainframe. Attachment of the subframe and the wheel suspension components to the mainframe can often involve complex, bulky, and expensive mounting features.

Vehicular frame assemblies and vehicles having support bracket


The present invention is a chemical reactor and method for catalytic chemical reactions having gas phase reactants. The chemical reactor has reactor microchannels for flow of at least one reactant and at least one product, and a catalyst material wherein the at least one reactant contacts the catalyst material and reacts to form the at least one product. The improvement, according to the present invention is: the catalyst material is on a porous material having a porosity that resists bulk flow therethrough and permits molecular diffusion therein. The porous material further has a length, a width and a thickness, the porous material defining at least a portion of one wall of a bulk flow path through which the at least one reactant passes.

The present invention is a chemical reactor and method for gas phase reactant catalytic reactions.
As used herein, the term xe2x80x9cmolecular diffusionxe2x80x9d is used in its classic sense of the transfer of mass based upon Brownian motion between adjacent layers of fluid in laminar, transition, or turbulent flow, and includes transfer of mass between adjacent layers of fluid that are stagnant.
As used herein, the term xe2x80x9cKnudsen diffusionxe2x80x9d means Knudsen flow, or free molecule flow, wherein the mean free path of the molecules is long compared to a characteristic dimension of the flow field, for example the pore size of a material through which the molecules are diffusing.
Many catalytic reactions begin with gas phase reactants, for example steam reforming, partial oxidation, water gas shift and others. However, equipment, specifically reactor volume is generally large because of mass and heat transfer limitations. Conventional reactors are operated with a gas hourly space velocity from about 1,000 to about 10,000 hrxe2x88x921. In other words, residence time is greater than 1 second because of the heat and mass transfer limitations.
These problems have been recognized and research is considering microchannel reactors because the microchannels have been shown to offer less resistance to heat and mass transfer thus creating the opportunity for dramatic reductions in process hardware volume. Several types of microchannel reactors have been described in the literature.
Franz et al., 1998 and Lowe et al., 1998 report applying a coating of the active catalyst (such as Pt, Ag, or other noble metal) directly to the microchannel wall. This approach has the disadvantage that the only usable surface area is that of the microchannel wall.
Weissmeier and Honicke, 1998a-b report creating a porous interface directly from the microchannel wall material onto which the catalyst is deposited. An aluminum wall was anodized to create the porous alumina interface that had an average pore diameter in the nanometer size range (permitting only Knudsen diffusion) and a thickness in the range of tens of microns. Disadvantages of this approach include that it is only applicable for aluminum, and limited surface area. The anodized walls formed a two-dimensional array of 700 identical microchannels.
Tonkovich/Zilka et al., 1998 reported packing catalytic powders directly within an array of parallel microchannels as a packed microbed. A disadvantage was a tendency to create relatively large pressure drops by forcing the fluid to flow through the packed microbed.
Tonkovich/Jimenez et al., 1998 reported placing a palladium catalyst supported on a metallic nickel foam within a cavity (more than an order of magnitude larger than a microchannel) and then sending the effluent to an array of microchannels to exchange heat. Again, a disadvantage was large pressure drop through the metal foam.
Hence, there is a need for a chemical reactor for catalytic reactions with fast kinetics that has a small reactor volume with a low pressure drop.
Franz, A. J., Quiram, D., Srinivasan, R., Hsing, I-M., Firebaugh, S. L., Jensen, K. F., and M. A. Schmidt, 1998, New Operating Regimes and Applications Feasible with Microreactors, Proceedings of the Second International Conference on Microreaction Technology, New Orleans, La., p 33-38.
Lowe, H., Ehrfeld, W., Gebauer, K., Golbig, K., Hausner, O., Haverkamp, V., Hessel, V., and Richter, Th., 1998, Microreactor Concepts for Heterogeneous Gas Phase Reactions, Proceedings of the Second International Conference of Microreaction Technology, March 1998, New Orleans, La., p. 63-74.
Tonkovich, A. Y., Zilka, J. L., Powell, M. R., and C. J. Call, 1998, The Catalytic Partial Oxidation of Methane in a Microchannel Chemical Reactor, Proceedings of the Second International Conference of Microreaction Technology, March 1998, New Orleans, La., p.45-53.
Tonkovich, A. Y., Jimenez, D. M., Zilka, J. L., LaMont, M., Wang, Y., and R. S. Wegeng, 1998, Microchannel Chemical Reactors for Fuel Processing, Proceedings of the Second International Conference of Microreaction Technology, March 1998, New Orleans, La., p.186-195.
Weissmeier, G., and Honicke, D., 1998a, Strategy for the Development of Micro Channel Reactors for Heterogeneously Catalyzed Reactions, Proceedings of the Second International Conference on Microreaction Technology, New Orleans, La., p. 24-32.
Weissmeier, G., and Honicke, D., 1998b, Microreaction Technology: Development of a microchannel reactor and its application in heterogeneously catalyzed hydrogenation, Proceedings of the Second International Conference on Microreaction Technology, New Orleans, La., p.152-153.
The present invention is a chemical reactor and method for catalytic chemical reactions having gas phase reactants. The chemical reactor has at least one reactor microchannel defining a bulk flow path through which at least one reactant passes, and at least one product, and a catalyst material wherein at least one reactant contacts the catalyst material and reacts to form at least one product. The improvement, according to the present invention is:
the catalyst material is in a porous structure having a porosity that permits molecular diffusion therein. The porous structure further has a length, a width and a thickness, the porous structure defining at least a portion of at least one wall of the at least one microchannel.
It is an object of the present invention to provide a chemical reactor for gas phase reactant catalytic reactions.
It is an object of the present invention to provide a method for gas phase reactant catalytic reactions.
A significant advantage of the present invention is the reduced pressure drop because the flow through the bulk flow path passes and contacts the porous structure but is not required to flow through the porous structure. Sufficient reaction still occurs because of the net flux through molecular diffusion into (gas phase reactant(s)) and out of (product(s)) the porous structure. Gas hourly space velocity is unexpectedly greater than 10,000 hrxe2x88x921 corresponding to residence time less than 1 second.
The subject matter of the present invention is particularly pointed out and distinctly claimed in the concluding portion of this specification. However, both the organization and method of operation, together with further advantages and objects thereof, may best be understood by reference to the following description taken in connection with accompanying drawings wherein like reference characters refer to like elements.

Chemical reactor and method for gas phase reactant catalytic reactions


Liquid flow through a conduit in a branch of a hydronic system provided with a balancing valve and, in series therewith, a metering valve to obtain a desired rate of flow in the branch is regulated by positioning immediately adjacent the metering valve in the conduit a pitot having an upstream side directed upstream against a direction of flow through the conduit and a downstream side directed downstream in the flow direction. The metering valve is settable in a plurality of predetermined positions each corresponding to a respective flow cross section through the conduit. A differential pressure meter is connected to both sides of the pitot, the one of the predetermined valve positions is selected that gives a near full-range reading on the meter, and the metering valve is set in this selected one position. The pressure-differential reading on the meter is monitored and the balancing valve is adjusted to regulate flow therethrough until the read pressure corresponds to a level itself corresponding to the desired rate of flow in the branch.

In a complex hydronic system, for instance a building heating system, hot water is pumped from a central boiler up a common riser from which it flows through a multiplicity of branch lines each including one or more terminals. Then the multiple streams are reunited in a common downpipe that leads back to the boiler. In such a system it is necessary to balance the flow in the individual branches. Thus each branch is provided with a balancing valve, which is nothing more than a lockable flow-control valve, that is adjusted until a predetermined flow, normally measured in gallons per minute, is obtained in the branch. In this manner a branch with particularly low resistance to flow does not get too much flow while another with relatively high resistance gets too little.
The standard method for adjusting flow in a branch, typical a radiator or coil, is fairly laborious. A typical-balancing valve (the Bell & Gossett ITT "Circuit Setter Plus," or the "CBV" system of Armstrong Pumps) has a graduated spindle that indicates the valve's flow cross section and its resistance to flow. Immediately upstream and downstream of the valve element of such a valve, the valve housing is provided with closable nipples that open internally into the valve. A differential pressure meter is first connected to these nipples. Then the balancing valve is adjusted to one of a multiplicity of graduated settings and the difference in pressure between the measuring locations upstream and downstream of the valve and the valve setting is noted. These readings are checked on a chart for the particular valve size to ascertain the flow rate in volume per units of time. The chart plots flow against differential pressure and has a large number of lines corresponding to the different settings so that the user checks the line for the setting being used, and determines for the pressure reading the flow. If the ascertained flow rate is not correct the valve is readjusted, a reading is taken again, and the appropriate line on the chart is consulted to get the new flow rate. Thus the balancing valve serves both for balancing and measuring flow. This process is repeated until the desired flow rate is obtained, and the valve is secured in the last position to lock in this flow rate, and a stop is set so that if the balancing valve is subsequently shut, for example for servicing of the associated radiator or chiller, it can be returned to the desired position, typically called the "memory" position.
Obviously this is an extremely laborious procedure involving making two separate connections to the line, and then adjusting the valve while frequently consulting a chart. The person doing this flow balancing must be fairly skilled, and even so the procedure can be very time consuming in a large building that can have hundreds of branches.
Another problem with the known system is that it is relatively inaccurate. The pressure-differential meter must normally be able to read pressure differentials over a relatively wide range. Like all such analog equipment, however, it is typically fairly accurate at one part, typically the high end, of its range but much less so toward the low end. Furthermore the valve positions must be very carefully monitored and the valves must be extremely accurately manufactured to ensure some close correspondence between the readings and the actual values. Thus if the flow in a line is relatively low the technician can be dealing with a combined inaccuracy of 20% or more.
Another problem with the known systems is that connecting the differential pressure meter to the branch is a laborious problem, entailing making two separate connections at two separate locations. There is frequently some leakage, and the taps provided for the differential pressure meter are themselves subject to leakage or vandalism.

Method of and valve for controlling flow in a hydronic system


A decorative snow guard assembly attachable to raised portions of a building surface such as a standing seam roof. The assembly includes a decorative snow brake plate integral with a base, means spanning between adjacent bases, and means connecting said spanning means to said bases.

With the increased use of sheet metal panels in building construction, there has been an increased need to address ways in which various building attachments can be interconnected with a metal panel surface. For example, in the case of metal roofs, there is often a need to mount/secure various types of equipments thereon. Specifically, in various climates, it may be desirable to position a snow retention device on a metal roof to control/inhibit/impede the movement of snow and/or ice down the pitch of the roof.
Sliding snow and/or ice from metal roofs can be hazardous to people, the surrounding landscape, property, and building components. For example, snow or ice sliding from a roof above an entryway may injure a passersby. Similarly, falling snow or ice can do damage to landscape features, such as shrubs, and property or building components, including automobiles or lower roofing portions. In addition, sliding snow or ice can shear off antennas, gutters or other components attached to a building roof or wall, thereby potentially causing a leak. The problem of sliding snow or ice is particularly experienced in connection with metal roofs, including raised seam roofs, (e.g., standing seam) where there is relatively little friction between the roof and the snow or ice.
Numerous snow guard devices have been developed and used in the prior art. However, there are a number of problems generally associated with one or more of the snow guard devices. For example, such devices may cause the roof to leak. Many of the prior art devices are attached to roof by a screw, nail, or other fastener which pierces the roofing surface. Such piercing of the roof can lead to undesirable leakage due to inadequate sealing or shearing of the fastener by the forces exerted thereon by sliding snow and/or ice. Alternate methods for attachment of snow guard devices to roofs such as adhesive bonding may fail to provide secure attachment and/or may be difficult to install on a sloped surface, particularly where the device is applied to a smooth non-porous roofing material such as metal.
Many known snow guard devices can also cause undesirable pinning of the roof materials. Metal roofing sheets are often designed to be movable so as to accommodate normal thermal expansions and contractions. Where snow guard devices in the prior art are attached to the roof by a screw, nail or the like, which pierces the roof surface and is anchored to an underlying structural member or deck, the design thermal movement characteristics of the roof can be compromised thereby adversely effecting the roof's performance. Based upon the foregoing, it is apparent there is a need for a mounting device for a decorative snow guard which may be positioned on a metal panel surface without adversely effecting its performance.


Systems and methods of semi-centralized energy storage and mobile power outflow for vehicle propulsion comprise at least one energy storage facility receiving energy via an electric grid and at least one mobile vehicle. The energy is generated at a first location, and the energy storage facility is at a second location different from the first location. The second location is closer to end users of the energy than the first location. The energy storage facility produces an energy storage medium at the second location and stores the energy from the first location at the second location in the energy storage medium. The energy storage medium comprises liquid air, liquid oxygen, liquid nitrogen, or a combination thereof. The mobile vehicle includes a prime mover and a cryogenic storage vessel and is configured to carry at least a portion of the energy storage medium in the cryogenic storage vessel and use power from the energy storage medium for mobile vehicle propulsion.

Storage of electric power allows low-value power produced "off-peak" (e.g., overnight) to be released during "peak" power demand periods (e.g., daytime) when the value of power is substantially higher and when power shortages are most likely to occur. This is true regardless of the source (e.g., coal-fired power, nuclear power, wind power, etc.) of the "original" power to be stored. Numerous important benefits result from multi-megawatt ("bulk") power storage methods, including using off-peak energy to provide more peak power, expanding the amount of higher-value peak power that can be delivered through existing transmission and distribution infrastructure, and transforming intermittent (unreliable) power sources such as wind and solar into "firm," "dispatchable" (reliable) sources.
Generally, electrical grids can only tolerate +/−10% intermittent renewable power as a percentage of the total power supply on the grid, because allowing significantly more intermittent power would cause the grid to become unstable/unreliable and may "crowd-out" the more reliable (and thus more valuable) power from baseload plants. This represents a "ceiling" for the market penetration of intermittent renewable power sources in the absence of a cost-effective, broadly deployable bulk power storage solutions such as the disclosed embodiments. In addition, smaller multi-megawatt energy storage systems (e.g., 1 MW to 50 MW) provide a means to add peak power capacity to constrained load pockets at high net efficiency while helping to upgrade (and, in effect, expand) power distribution systems, especially in areas where few other options exist and/or where upgrades are costly.
There are many benefits of bulk power storage to the electrical grid, including but not limited to the ability to buy electric energy low (off-peak) and sell high (peak), greater electric supply capacity, elimination of the need for "peaker" power plants, reduction of transmission capacity requirements and congestion, better electric service reliability and power quality, reduction in electric bills, and firming of renewables capacity. Not all of the above power storage values apply for each deployment of a storage asset, because the individual values that can be garnered are case-specific. However, as a general rule, the closer the power "release" of a power storage system is to the load, the more benefit it will bring to the overall electrical grid, including but not limited to, utilities, ISOs and rate-payers. Thus, systems such as the disclosed embodiments whose power release can occur close to the power load are more valuable than those that can only be deployed far "upstream" on the grid.
Despite the many advantages of bulk power storage identified above, there are very few market-ready power storage options in the lower commercial scale range, e.g., 1 MW to 50 MW. Battery technologies are generally limited to a few MW/MWH of capacity (under 5 MW/10 MWH per day) and have other drawbacks including disposal issues at the end of the life of the batteries and limited discharge durations (generally less than 2 hours per day). There are several available methods for large, utility-scale (greater than 100 MW) power storage, including Compressed Air Energy Storage (CAES), pumped hydro, and liquid air energy storage (LAES) such as Vandor's Power Storage (VPS) Cycle. CAES is severely limited by the need for caverns and/or other underground geological formations that are required to contain the compressed air. Also, CAES does not deliver a consistent amount of power output; rather, the amount of power generated declines with each hour of power release, as the pressure of the compressed air in the cavern decreases throughout the daily cycle. Pumped hydro is also limited by geography and geology, requiring two large "lakes" to be separated by a significant height differential between the two reservoirs, with a dam and water-turbines between them.
VPS is much more flexible because it is not constrained by geography/geology and can be placed near the end user of the power, i.e., at the load, thus enhancing the deployment's value. However, in order to increase the total number of possible deployment sites, the basic principles of VPS, and the methods and systems embodied in the previous patents, need to be enhanced to allow for the technology to be deployable not only at utility scales, (which can be characterized as approximately 50 MW/400 MWH and larger), but also at smaller, "commercial scales," which will typically be as small as about 1 MW, but could be even smaller, i.e., in the kW scale. A network of widely deployed, smaller-than utility-scale, power storage facilities will constitute a "distributed power storage" system. Such a system offers many benefits, including flexibility as to how the inflow to storage occurs on the grid and as to how the outflow from storage is sent out to power users.
The various pumped hydro, and the two CAES deployments now operating worldwide, are the largest-scale examples of power storage, where each deployment can send out hundreds of megawatts of power. At the other end of the scale, proposals have been put forth for using the power storage capacity of the batteries in all-electric cars as a "crowd source" distributed power network where each car provides only a few kW of power output (when it is plugged in to a two-way connection to the grid), but where the cumulative effect of many such plugged in cars may be significant. Thus, there is a need for an energy storage system that fits within the gap that exists between those two extremes, and which gap is not now served by commercially viable technologies.
In addition to the power storage issues outlined briefly above, the power distribution network faces another set of related challenges that are best solved by "distributed power generation" systems and methods. For example, if the power distribution network is to be more reliable, it needs to be restructured. In particular, there is a need for an energy production and distribution system that relies less on large-scale, centralized power plants located far from their customers, where an increasingly congested and vulnerable (to natural and man-made events) grid connects large generating facilities with a customer base. Instead, there is a need for power generation that can be employed at lower capital cost and more locally, near customers, providing for redundancy, shorter travel times/distances on the grid (reducing line losses and congestion), and allowing for more competition in the market and at a pricing structure that reflects the multiple options for localized (distributed) power production. The smaller the economic and technical "threshold" for distributed power production systems the more widely they can be deployed, the more "distributed" the deployments, the more such a network can avoid grid congestion, and the less vulnerable the grid becomes to unplanned natural and man-made outages i.e., by relying on numerous widespread generation sources instead of just one (or several) large, centralized power plants).
Accordingly, there is a need for power storage systems and methods, which can transform low-value, off-peak power into high-value peak power and which can make intermittent power sources "firm." There is a further need for power storage and distributed generation systems and methods that can be widely deployed at smaller commercial scales. Finally, There is also a need for power storage and distributed generation systems and methods, which can be located close to power consumers (i.e., load centers).

Systems and methods of semi-centralized power storage and power production for multi-directional smart grid and other applications