Optimizing E. coli-based membrane protein production using Lemo21(DE3) and GFP-fusions

Methods Mol Biol. 2013:1033:381-400. doi: 10.1007/978-1-62703-487-6_24.

Abstract

Optimizing the conditions for the overexpression of membrane proteins in E. coli and their subsequent purification is usually a laborious and time-consuming process. Combining the Lemo21(DE3) strain, which conveniently allows to identify the optimal expression intensity of a membrane protein using only one strain, and membrane proteins C-terminally fused to Green Fluorescent Protein (GFP) greatly facilitates the production of high-quality membrane protein material for functional and structural studies.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bioreactors
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism*
  • Fermentation
  • Gene Expression Regulation, Bacterial
  • Genetic Vectors / genetics
  • Green Fluorescent Proteins / biosynthesis*
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics
  • Membrane Proteins / biosynthesis*
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Transformation, Bacterial

Substances

  • Membrane Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins