VECTORS FOR CU2+-INDUCIBLE PRODUCTION OF GLUTATHIONE S-TRANSFERASE-FUSION PROTEINS FOR SINGLE-STEP PURIFICATION FROM YEAST

被引：24

作者：

WARD, AC ^{[1
]}

CASTELLI, LA ^{[1
]}

MACREADIE, IG ^{[1
]}

AZAD, AA ^{[1
]}

机构：

[1] UNIV MELBOURNE, RUSSELL GRIMWADE SCH BIOCHEM, PARKVILLE, VIC 3052, AUSTRALIA

来源：

YEAST | 1994年 / 10卷 / 04期

关键词：

SACCHAROMYCES CEREVISIAE; CUP1; CU2+-REGULATED; YEAST EXPRESSION; AFFINITY PURIFICATION; GLUTATHIONE S-TRANSFERASE; METALLOTHIONEIN;

D O I：

10.1002/yea.320100403

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe six new yeast episomal vectors which encode glutathione S-transferase (GST) affinity tags. These allow for the production of GST-fusion proteins in Saccharomyces cerevisiae under the control of the CUP1 promoter. Affinity chromatography with glutathione-Sepharose permits convenient purification of the fusion protein from a yeast lysate. The presence of a protease cleavage site facilitates subsequent removal of the GST tag. The expression and single-step purification of both GST and a functional GST-metallothionein fusion from yeast are shown as an example of the application of these vectors.

引用

页码：441 / 449

页数：9

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