大肠杆菌中膜蛋白的生产。

Membrane Protein Production in Escherichia coli.

机构信息

Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.

出版信息

Methods Mol Biol. 2020;2127:13-27. doi: 10.1007/978-1-0716-0373-4_2.

Abstract

Escherichia coli is the workhorse of the structural biology lab. In addition to routine cloning and molecular biology, E. coli can be used as a factory for the production of recombinant membrane proteins. Purification of homogeneous samples of membrane protein expressed in E. coli is a significant bottleneck for researchers, and the protocol we present here for the overexpression and purification of membrane proteins in E. coli will provide a solid basis to develop lab- and protein-specific protocols for your membrane protein of interest. We additionally provide extensive notes on the purification process, as well as the theory surrounding principles of purification.

摘要

大肠杆菌是结构生物学实验室的主力。除了常规的克隆和分子生物学，大肠杆菌还可以作为生产重组膜蛋白的工厂。在大肠杆菌中表达的同质膜蛋白的纯化是研究人员的一个重要瓶颈，我们在这里提出的用于在大肠杆菌中过表达和纯化膜蛋白的方案将为您感兴趣的膜蛋白提供实验室和蛋白特异性方案的坚实基础。我们还提供了关于纯化过程的广泛说明，以及围绕纯化原理的理论。