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一种用于细菌表达蛋白的新型同位素标记方案。

A novel isotope labeling protocol for bacterially expressed proteins.

作者信息

Reilly D, Fairbrother W J

机构信息

Department of Cell Culture, Genentech Inc., South San Francisco, CA 94080-4990.

出版信息

J Biomol NMR. 1994 May;4(3):459-62. doi: 10.1007/BF00179352.

Abstract

A novel protocol for isotopically labeling bacterially expressed proteins is presented. This method circumvents problems related to poor cell growth, commonly associated with the use of minimal labeled media, and problems with protein induction encountered, less commonly, when using enriched labeled media. The method involves initially growing the bacterial cells to high optical density in a commercially available enriched labeled medium. Following a suitable growth period, the cells are transferred to a different (minimal) labeled medium, appropriate for induction. The method is demonstrated using the protein melanoma growth stimulating activity (MGSA).

摘要

提出了一种用于对细菌表达的蛋白质进行同位素标记的新方案。该方法规避了与细胞生长不良相关的问题,这种问题通常与使用最低限度标记的培养基有关,以及在使用富集标记的培养基时较少遇到的蛋白质诱导问题。该方法包括首先在市售的富集标记培养基中将细菌细胞培养至高光密度。经过适当的生长阶段后,将细胞转移到适合诱导的不同(最低限度)标记培养基中。使用蛋白质黑素瘤生长刺激活性(MGSA)对该方法进行了验证。

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