Document Detail


Vectors for the inducible overexpression of glutathione S-transferase fusion proteins in yeast.
MedLine Citation:
PMID:  8368005     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
D A Mitchell; T K Marshall; R J Deschenes
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Yeast (Chichester, England)     Volume:  9     ISSN:  0749-503X     ISO Abbreviation:  Yeast     Publication Date:  1993 Jul 
Date Detail:
Created Date:  1993-10-06     Completed Date:  1993-10-06     Revised Date:  2009-07-22    
Medline Journal Info:
Nlm Unique ID:  8607637     Medline TA:  Yeast     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  715-22     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, University of Iowa, Iowa City 52242.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Fungal Proteins / genetics
Galactose
Gene Expression / genetics
Genes, Fungal / genetics*
Genetic Vectors / genetics*
Glutathione Transferase / biosynthesis,  genetics*
Molecular Sequence Data
Plasmids
Protein Processing, Post-Translational
Recombinant Fusion Proteins / biosynthesis,  genetics*
Saccharomyces cerevisiae / enzymology*,  genetics
Saccharomyces cerevisiae Proteins*
ras Proteins*
Grant Support
ID/Acronym/Agency:
CA50211/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Fungal Proteins; 0/Recombinant Fusion Proteins; 0/Saccharomyces cerevisiae Proteins; 26566-61-0/Galactose; EC 2.5.1.18/Glutathione Transferase; EC 3.6.5.2/RAS2 protein, S cerevisiae; EC 3.6.5.2/ras Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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