( Epitope-Tagging Vectors for Functi...)Epitope-Tagging Vectors for Functional Analysis in Yeast

Regulation of gene expression in yeast

Quinn Lu Tanya Hosfield
Stratagene

Stratagene has developed a series of epitope-tagging vectors to express and functionally analyze eukaryotic genes in the budding yeast Saccharomyces cerevisiae. Each of these pESC vectors contains one of four different yeast selective markers in the same vector backbone, which allows expression and epitope-tagging analysis of two different genes in a single yeast cell.

Characterizing the structure and function of every eukaryotic gene is a necessary task in the post genome era. Results of the yeast genome sequencing project indicate that S. cerevisiae shows homologous sequences for one-third of the human disease genes discovered to date.¹ Hence, the yeast makes an attractive model for studying these genes.

We constructed a series of epitope-tagging vectors to facilitate the in vivo study of eukaryotic genes in S. cerevisiae. With these vectors, any cloned gene can be introduced into yeast under the control of a repressible promoter, two different genes can be coexpressed in yeast, protein-protein interactions can be confirmed by immunoprecipitation analysis, and protein localization can be studied. Together with the use of other vectors, the pESC vectors can be used to characterize functional domains of an essential gene and genetically screen for suppressors and inhibitors of certain types of genes in yeast.

pESC Vectors

Figure 1

The pESC vectors (Figure 1) are specifically designed to express high levels of eukaryotic proteins in S. cerevisiae. They are derived from the pRS400 series vectors.² Each pESC vector contains the following seque
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