Vectors for Expressing T7 Epitope- and His 6 Affinity-Tagged Fusion Proteins in S. cerevisiae

We have constructed a series of vectors (YGALSETs) for the expression of epitope- and affinity-tagged fusion proteins in yeast cells using the regulated GAL10 promoter. Fusion proteins produced from YGALSET plasmids include a leader peptide at the N terminus that encodes both a T7 gene 10 epitope tag and a His6 affinity tag. The YGALSET vector series includes centromere plasmids for low-copy plasmid maintenance and 2 micron episomal plasmids for multicopy plasmid maintenance and four different selectable markers: TRP1, URA3, LEU2 and HIS3. We also provide a convenient approach for transferring cloned genes from a bacterial expression vector into YGALSET vectors by in vivo recombination and a rapid method to screen directly for clones that express the fusion protein of interest.