Mitchell D A, Marshall T K, Deschenes R J
Department of Biochemistry, University of Iowa, Iowa City 52242.
Yeast. 1993 Jul;9(7):715-22. doi: 10.1002/yea.320090705.
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.
一种快速便捷的蛋白质纯化方法是构建与谷胱甘肽S-转移酶（GST）的融合蛋白（Smith和Johnson，《基因》67卷，31 - 40页，1988年）。在本报告中，我们描述了两种用于在酿酒酵母中条件性表达GST融合蛋白的载体。亲本质粒基于先前描述的高拷贝、半乳糖诱导型穿梭载体（Baldari等人，《欧洲分子生物学组织杂志》6卷，229 - 243页，1987年）。我们通过构建GST与酵母RAS2基因之间的融合蛋白证明了该系统的用途。GST-Ras2融合蛋白会经历Ras2p定位于膜所需的翻译后修饰。这些载体为表达和纯化需要翻译后修饰的真核蛋白提供了一个有用的系统。
