Abstract
Functional and structural studies on membrane proteins are limited by the difficulty to produce them in large amount and in a functional state. In this review, we provide protocols to achieve high-level expression of membrane proteins in Escherichia coli. The T7 RNA polymerase-based expression system is presented in detail and protocols to assess and improve its efficiency are discussed. Protocols to isolate either membrane or inclusion bodies and to perform an initial qualitative test to assess the solubility of the recombinant protein are also included.
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Acknowledgments
This work was supported by the Centre National de la Recherche Scientifique, INSERM, and by the “Initiative d'Excellence” program from the French State (Grant “DYNAMO,” ANR-11-LABEX-0011-01). FA is supported by a DYNAMO PhD fellowship.
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Angius, F., Ilioaia, O., Uzan, M., Miroux, B. (2016). Membrane Protein Production in Escherichia coli: Protocols and Rules. In: Mus-Veteau, I. (eds) Heterologous Expression of Membrane Proteins. Methods in Molecular Biology, vol 1432. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3637-3_3
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DOI: https://doi.org/10.1007/978-1-4939-3637-3_3
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