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.
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Acknowledgment
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.
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McIlwain, B.C., Kermani, A.A. (2020). Membrane Protein Production in Escherichia coli. In: Perez, C., Maier, T. (eds) Expression, Purification, and Structural Biology of Membrane Proteins. Methods in Molecular Biology, vol 2127. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0373-4_2
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DOI: https://doi.org/10.1007/978-1-0716-0373-4_2
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