Membrane Protein Production in Escherichia coli
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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.
Key wordsMembrane protein E. coli Crystallography Ion channel Transporter.
We would like to thank Randy Stockbridge as well as other members of the Stockbridge lab for helpful suggestions, guidance, and feedback on this chapter.
- 24.Bird LE, Rada H, Verma A, Gasper R, Birch J, Jennions M, Lӧwe J, Moraes I, Owens RJ (2015) Green fluorescent protein-based expression screening of membrane proteins in Escherichia coli. J Vis Exp 95:e52357Google Scholar