Format

Send to

Choose Destination
Methods Mol Biol. 2016;1432:37-52. doi: 10.1007/978-1-4939-3637-3_3.

Membrane Protein Production in Escherichia coli: Protocols and Rules.

Author information

1
Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, Institut de Biologie Physico-Chimique, CNRS, Univ Paris Diderot, Sorbonne Paris Cité, PSL research university, Paris, France.
2
Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, Institut de Biologie Physico-Chimique, CNRS, Univ Paris Diderot, Sorbonne Paris Cité, PSL research university, Paris, France. Bruno.Miroux@ibpc.fr.
3
Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, UMR7099 CNRS IBPC, 13 rue Pierre et Marie Curie, 75005, Paris, France. Bruno.Miroux@ibpc.fr.

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.

KEYWORDS:

E. coli; Production of recombinant proteins; T7 RNA polymerase

PMID:
27485328
DOI:
10.1007/978-1-4939-3637-3_3
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center