Gene Expression in Yeast

Abstract

In the past few decades expression of foreign genes in yeast has not only been a very powerful tool for analysing and understanding protein function within the framework of scientific investigations, it also has become a major economic factor of the biotechnological industries. The article presented here deals with well‐established methods and strategies for ensuing reliable transfer of foreign genes and sustained protein production in yeast.

Keywords: yeast; heterologous expression; YIp (yeast‐integrating plasmid) vectors; YRp (yeast‐replicating plasmid) vectors; YCp (yeast‐centromere plasmid) vectors; YEp (yeast‐episomal plasmid) vectors

Figure 1.

A generalized presentation of a YEp‐type shuttle vector. The plasmid shown combines all functional components necessary for successful transformation, maintenance of the plasmid, and enhanced transcription of the foreign gene. The sequences derived from the bacterial pBR322 plasmid contain a bacterial origin of replication and the bla and tet genes that confer resistance to ampicillin and tetracycline. Therefore, bacteria bearing such shuttle vectors will grow in selective media containing either of the two antibiotics. Propagation of the plasmid in yeast is ensured by including the origin of replication derived from the 2‐μm‐circle yeast plasmid. For maintenance in auxotrophic yeast strains, the plasmid contains LEU2, TRP1 or other genes as selectable markers. Finally, promoter and terminator sequences, separated by a multiple cloning site sequence that can be used for introducing the foreign DNA into the plasmid, complete the overall construction.

Figure 2.

DNA transfer by a YIp vector. In contrast to YEp vectors, YIp vectors do not contain the origin of replication from the 2‐μm‐circle yeast plasmid. Their propagation is ascertained through integration within specific loci of the yeast chromosome by means of homologous recombination. The expression of the integrated DNA fragment depends on the promoter of the yeast gene locus that was targeted by the YIp vector.

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References

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Further Reading

Agnan J, Korch C and Selitrennikoff C (1997) Cloning heterologous genes: problems and approaches. Fungal Genetics and Biology 21: 292–301.

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Maya D, Quintero MJ, de la Cruz Munoz‐Centeno M and Chavez S (2008) Systems for applied gene control in Saccharomyces cerevisiae. Biotechnology Letters 30: 979–987.

Trueman LJ (1995) Heterologous expression in yeast. Methods in Molecular Biology 49: 341–354.

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How to Cite close
Scheiner‐Bobis, Georgios(Mar 2009) Gene Expression in Yeast. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002666.pub2]