Gene Expression in Yeast


Yeast strains such as Saccharomyces cerevisiae, Pichia pastoris, Hansenula polymorpha, Kluyveromyces lactis and others constitute a significant alternative to prokaryotic or higher eukaryotic expression systems. As single‐cell organisms, they offer many of the advantages of prokaryotic expression systems such as inexpensive propagation and easy molecular manipulation. Because they are eukaryotes, however, yeast cells possess all organelles and mechanisms required for the expression of membrane‐directed, secretory and glycosylated proteins. 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 has also 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.

Key Concepts

  • Plasmids are circular, double‐stranded, extrachromosomal, autonomously replicating DNA molecules found in bacteria and archaea. Plasmids frequently provide antibiotic resistance to their host cells. As they are replicons with a single origin of replication (ORI), plasmids are often modified so that they can be used as vectors.
  • Vectors are vehicles used for the transfer of DNA into a recipient cell. Thus, a vector can be even an animal, such as the yellow fever mosquito (Aedes aegypti, formerly Stegomyia aegypti), an insect that propagates the yellow fever virus or the Zika virus. In molecular biology, the major types of vectors are modified plasmids, cosmids, phages, viral particles or artificial chromosomes.
  • Shuttle vectors are vectors that can be propagated in two different hosts (for example, in the prokaryotic Escherichia coli and also in the eukaryotic Saccharomyces cerevisiae).
  • Heterologous expression refers to the expression of a gene in an organism that does not naturally express the same gene. In contrast, homologous expression refers to the expression of host's own genes.
  • ORI (origin of replication) is the DNA segment where DNA replication initiates. Most circular chromosomes of prokaryotes carry a single ORI. Linear eukaryotic chromosomes might have up to 1000 ORIs.

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 generalised 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 (deoxyribonucleic acid) 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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Further Reading

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Mokdad‐Gargouri R, Abdelmoula‐Soussi S, Hadiji‐Abbès N, et al. (2012) Yeasts as a tool for heterologous gene expression. Methods in Molecular Biology 824: 359–370.

Matsuyama A and Yoshida M (2012) Heterologous gene expression by chromosomal integration in fission yeast. Methods in Molecular Biology 824: 433–450.

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Scheiner‐Bobis, Georgios(Jan 2018) Gene Expression in Yeast. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0002666.pub3]