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Membrane Protein Expression in Cell-Free Systems

  • Birgit Schneider
  • Friederike Junge
  • Vladimir A. Shirokov
  • Florian Durst
  • Daniel Schwarz
  • Volker Dötsch
  • Frank BernhardEmail author
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 601)

Abstract

Cell-free expression has emerged as a promising tool for the fast and efficient production of membrane proteins. The rapidly growing number of successfully produced targets in combination with the continuous development of new applications significantly promotes the distribution of this technology. Membrane protein synthesis by cell-free expression does not appear to be restricted by origin, size or topology of the target, and its global application is therefore a highly valuable characteristic. The technology is relatively fast to establish in standard biochemical labs, and it does not require expensive equipment. Moreover, it enables the production of membrane proteins in completely new modes, like the direct translation into detergent micelles, which is not possible with any other expression system. In this protocol, we focus on the currently most efficient cell-free expression system for membrane proteins based on Escherichia coli extracts.

Key words

Cell extracts cell-free expression system coupled transcription/translation detergents integral membrane protein solubilization 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Birgit Schneider
    • 1
  • Friederike Junge
    • 1
  • Vladimir A. Shirokov
    • 2
  • Florian Durst
    • 1
  • Daniel Schwarz
    • 1
  • Volker Dötsch
    • 1
  • Frank Bernhard
    • 1
    Email author
  1. 1.Centre for Biomolecular Magnetic Resonance, University of Frankfurt/Main, Institute for Biophysical ChemistryFrankfurt/MainGermany
  2. 2.Institute of Protein Research, Russian Academy of SciencesPushchinoRussia

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