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Vectors for the inducible overexpression of glutathione S‐transferase fusion proteins in yeast
Author(s) -
Mitchell David A.,
Marshall Tricia K.,
Deschenes Robert J.
Publication year - 1993
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320090705
Subject(s) - shuttle vector , biology , fusion protein , glutathione s transferase , yeast , saccharomyces cerevisiae , plasmid , fusion gene , expression vector , biochemistry , transferase , tandem affinity purification , gene , microbiology and biotechnology , glutathione , recombinant dna , vector (molecular biology) , affinity chromatography , enzyme
A rapid and convenient method of protein purification involves creating a fusion protein with glutathione S‐transferase (GST) (Smith and Johnson, Gene 67 , 31–40, 1988). In this report, we describe two vectors for the conditional expression of GST fusions in Saccharomyces cerevisiae . The parent plasmid is based on a high‐copy, galactose‐inducible shuttle vector previously described (Baldari et al., EMBO J. 6 , 229–243, 1987). We have demonstrated the use of this system by creating fusions between GST and the yeast RAS2 gene. GST‐Ras2 fusion proteins undergo the post‐translational modifications required for Ras2p to become membrane localized. These vectors provide a useful system for the expression an dpurification of eukaryotic proteins requiring post‐translational modification.