Membrane Protein Production in Escherichia coli: Protocols and Rules

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

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.

Publication types

  • Review
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • DNA-Directed RNA Polymerases / metabolism*
  • Escherichia coli / genetics
  • Escherichia coli / growth & development*
  • Guidelines as Topic
  • Membrane Proteins / biosynthesis*
  • Membrane Proteins / genetics
  • Protein Engineering
  • Recombinant Proteins / biosynthesis
  • Solubility
  • Viral Proteins / metabolism*

Substances

  • Membrane Proteins
  • Recombinant Proteins
  • Viral Proteins
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases