in non-dividing cells, allowing for stable transfer in a large range of host cell types. Lipofection In lipofection, the gene is injected with the help of liposomes. The DNA sequence is encapsulated in a liposome with the same composition as the cell membrane. This method allows it to directly fuse with the membrane, or be endocytosed, which then releases the DNA into the cell. Lipofection is often used because it works with many different cell types, is highly reproducible, and is a fast method for both stable and transient expression. Host Systems Genes are subjected to heterologous expression often to study specific protein interactions. E. coli, yeast (S. cerevisiae, P. pastoris), immortalized mammalian cells, and amphibian oocytes (i.e. unfertilized eggs) are commonly for studies that require heterologous expression. In choosing a particular system, economic and qualitative aspects have to be considered. Prokaryotic expression is widely used in recombinant DNA technology to form easily manipulated proteins by well-known genetic methods with a low costing medium. Some limitations include intracellular accumulation of heterologous proteins, improper folding of the peptide, lack of post-transcriptional modifications, the potential for product degradation due to traces of protease impurities, and production of endotoxin. Prokaryotic and eukaryotic systems, most commonly bacteria, yeast, insects, and mammalian cells, and occasionally amphibians, fungi, and protists are used for studies that require heterologous expression. Bacteria, especially E. coli, yeast (S. cerevisiae, P. pastoris), insects, and amphibian (oocyte) cells have been used as effective hosts for expressing foreign proteins. Generally, prokaryotes are easier to work with and better understood and are often the preferable host system. It is widely used in recombinant DNA technology to form easily manipulated proteins by well-known genetic methods with a low costing medium. For membrane proteins though, researchers have observed that mammalian cells are more effective. This is because there is a lack of post-transcriptional modifications in prokaryotic systems. Limitations include intracellular accumulation of heterologous proteins, improper folding of the peptide, the potential for product degradation due to trace of protease impurities, and production of endotoxin. Escherichia Coli A popular system utilized is Escherichia coli because of its rapid growth rate (~20–30 minutes), capacity for continuous fermentation and relatively low cost. Additionally, yeast has the capacity to express a high relative volume of heterologous protein. Specifically, up to 30% of proteins produced in yeast can be the heterologous gene product. There also are safe strains of E. coli