Vectors for the inducible overexpression of glutathione S-transferase fusion proteins in yeast

Yeast. 1993 Jul;9(7):715-22. doi: 10.1002/yea.320090705.

Abstract

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 and purification of eukaryotic proteins requiring post-translational modification.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Fungal Proteins / genetics
  • Galactose
  • Gene Expression / genetics
  • Genes, Fungal / genetics*
  • Genetic Vectors / genetics*
  • Glutathione Transferase / biosynthesis
  • Glutathione Transferase / genetics*
  • Molecular Sequence Data
  • Plasmids
  • Protein Processing, Post-Translational
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics*
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins*
  • ras Proteins*

Substances

  • Fungal Proteins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • Glutathione Transferase
  • RAS2 protein, S cerevisiae
  • ras Proteins
  • Galactose