DOI: 10.1002/yea.320090705. Vectors for the inducible overexpression of glutathione S‐transferase fusion proteins in yeast

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

10.1002/yea.320090705

Crossref journal-article

Wiley

Yeast (311)

Abstract

AbstractA 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.

Bibliography

Mitchell, D. A., Marshall, T. K., & Deschenes, R. J. (1993). Vectors for the inducible overexpression of glutathione Sâtransferase fusion proteins in yeast. Yeast, 9(7), 715â722. Portico.

Authors 3

David A. Mitchell (first)
Tricia K. Marshall (additional)
Robert J. Deschenes (additional)

References 19 Referenced 248

{'key': 'e_1_2_1_2_1', 'volume-title': 'Current Protocols in Molecular Biology', 'author': 'Ausubel F. M.', 'year': '1991'} / Current Protocols in Molecular Biology by Ausubel F. M. (1991)
10.1002/j.1460-2075.1987.tb04743.x
10.1016/0003-2697(76)90527-3
10.1093/genetics/93.1.51 / Genetics / Selection of lys2 mutants of the yeast Saccharomyces cerevisiae by the utilization of alpha aminoadipate by Chattoo B. B. (1979)
10.1146/annurev.bi.61.070192.002035
{'key': 'e_1_2_1_7_1', 'first-page': '331', 'article-title': 'Isoprenoid modification and plasma membrane association: Critical factors for ras oncogenicity', 'volume': '3', 'author': 'Der C. J.', 'year': '1991', 'journal-title': 'Cancer Cells'} / Cancer Cells / Isoprenoid modification and plasma membrane association: Critical factors for ras oncogenicity by Der C. J. (1991)
10.1128/MCB.7.7.2344
Frangioni J. V.andNeel B. G.(1993). Solubilization and purification of enzymatically active glutathione S‐transferase (pGEX) fusion proteins.Anal. Biochem.in press. (10.1006/abio.1993.1170)
10.1016/S0021-9258(18)98789-2 / J. Biol. Chem. / Cloning and expression of a yeast protein tyrosine phosphatase by Guan K. (1991)
10.1016/0003-2697(91)90534-Z
10.1073/pnas.78.4.2199
10.1016/0003-2697(92)90108-J
10.1128/jb.153.1.163-168.1983
10.1016/0092-8674(85)90304-6
10.1016/S0021-9258(18)35774-0 / J. Biol. Chem. / Ras (CXXX) and Rab (CC/CXC) prenylation signal sequences are unique and functionally distinct by Khosavi‐Far R. (1992)
10.1038/227680a0
{'key': 'e_1_2_1_18_1', 'volume-title': 'Laboratory Course Manual: Methods in Yeast Genetics', 'author': 'Sherman F.', 'year': '1986'} / Laboratory Course Manual: Methods in Yeast Genetics by Sherman F. (1986)
10.1016/0378-1119(88)90005-4
10.1016/0092-8674(85)90305-8

Dates

Type	When
Created	20 years, 2 months ago (May 28, 2005, 7:44 p.m.)
Deposited	1 year, 9 months ago (Oct. 24, 2023, 10:29 p.m.)
Indexed	1 week, 4 days ago (Aug. 12, 2025, 6:24 p.m.)
Issued	32 years, 1 month ago (July 1, 1993)
Published	32 years, 1 month ago (July 1, 1993)
Published Online	21 years, 6 months ago (Jan. 29, 2004)
Published Print	32 years, 1 month ago (July 1, 1993)

Funders 0

None

BibTeX

@article{Mitchell_1993, title={Vectors for the inducible overexpression of glutathione S‐transferase fusion proteins in yeast}, volume={9}, ISSN={1097-0061}, url={http://dx.doi.org/10.1002/yea.320090705}, DOI={10.1002/yea.320090705}, number={7}, journal={Yeast}, publisher={Wiley}, author={Mitchell, David A. and Marshall, Tricia K. and Deschenes, Robert J.}, year={1993}, month=jul, pages={715–722} }

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