Open AccessVectors for Expressing T7 Epitope- and His6 Affinity-Tagged Fusion Proteins in S. cerevisiae
Author(s) -
Shinichiro Enomoto,
Guanghui Chen,
Judith Berman
Publication year - 1998
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/98245st01
Subject(s) - ura3 , plasmid , fusion protein , biology , epitope , microbiology and biotechnology , expression vector , multiple cloning site , gene , selectable marker , vector (molecular biology) , fusion gene , chimeric gene , genetics , recombinant dna , gene expression , antigen
We have constructed a series of vectors (YGALSETs) for the expression of epitopeand 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 His 6 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.