Vectors for expressing T7 epitope- and His6 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.
Enomoto
S
S
University of Minnesota, St. Paul, USA.
Chen
G
G
Berman
J
J
D000345:Affinity Labels; D017123:Bacteriophage T7 / Q000235:genetics*; D002213:Capsid / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D000939:Epitopes / Q000235:genetics*; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000378:metabolism*; D006639:Histidine / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000138:chemical synthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D000345:Affinity Labels; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D006639:Histidine