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Yeast

Genetic Transformation

Finally, one of the most useful properties of yeast for genetic studies is the ease with which DNA can be introduced into them, in a process called transformation. The introduced DNA can be maintained on self-replicating, circular strands of DNA called plasmids, or it can integrate into the yeast genome. Most importantly, integration usually occurs by a process called homologous recombination, whereby the introduced DNA replaces chromosomal DNA that contains the same sequence. This process permits scientists to readily mutate any yeast gene and replace the native gene in the cells with the mutated version. Since yeast can be grown as haploids, the phenotypic changes caused by the introduced gene can be readily identified. In addition, the function of a cloned piece of DNA (e.g., a gene) can be identified by transforming yeast in which the DNA is carried on a circular plasmid. The introduced gene may either functionally replace a defective gene or cause a phenotypic defect in the cells indicating a function for that gene.

The ability to complement yeast defects with cloned pieces of DNA has been extended to mammalian genes. Recognizing that some genes have similar sequences and functions in both mammals and yeast, scientists sometimes use yeast as a tool to identify the functions of mammalian genes. Not many mammalian genes can directly substitute for a yeast gene, however. More frequently, scientists study the yeast gene itself to understand how its protein functions in the cell. The knowledge gained can often lead to an understanding of how similar genes might function in mammals. Now that the yeast genome has been completely sequenced and the results have been deposited in a public databank for all to use, rapid progress is being made in identifying all yeast genes and their functions.

An important method for studying mammalian genes in yeast is called the two-hybrid system. This system is used to determine if two proteins functionally interact with each other. Both genes are cloned into yeast plasmids and transformed into the cells. A special detection system is used that is active only when both cloned proteins physically contact each other in the cell. When that happens, scientist can identify which proteins need to interact with each other in order to function.

Yeast are also being used in the laboratory and commercial production of important nonyeast proteins. Foreign genes are transformed into yeast and, after transcription and translation, the foreign proteins can be isolated. Because of the ease of growing large quantities of cells, yeast can produce a large amount of the protein. While similar protein production can be performed by bacteria, eukaryotic proteins often do not function when made in bacteria. This is because most eukaryotic proteins are normally altered after translation by the addition of short sugar chains, and these modifications are often required for proper function, but bacteria do not carry out these necessary post-translational modifications. Yeast, however, does permit these modifications, and is thus more likely to produce a functional protein.

Suzanne Bradshaw

Bibliography

Sherman, Fred. "Getting Started with Yeast." In Methods in Enzymology, vol. 194, Christine Guthrie and Gerald R. Fink, eds. New York: Academic Press, 1991.

Watson, James D., Michael Gilman, Jan Witowski, and Mark Zoller. Recombinant DNA. New York: Scientific American Books, 1992.

Additional topics

Medicine EncyclopediaGenetics in Medicine - Part 4Yeast - Yeast Genetics, Genetic Transformation