4.6 Article

Efficient production and purification of functional bacteriorhodopsin with a wheat-germ cell-free system and a combination of Fos-choline and CHAPS detergents

Journal

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2010.08.119

Keywords

Bacteriorhodopsin; Cell-free expression; Membrane protein; Micelle

Funding

  1. Ministry of Education, Culture, Sports, Science, and Technology of Japan [20053016]
  2. Grants-in-Aid for Scientific Research [20053016] Funding Source: KAKEN

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Cell-free translation is one potential approach to the production of functional transmembrane proteins. We have now examined various detergents as supplements to a wheat-germ cell-free system in order to optimize the production and subsequent purification of a functional model transmembrane protein, bacteriorhodopsin. We found that Fos-choline and CHAPS detergents counteracted each other's inhibitory effects on cell-free translation activity and thereby allowed the efficient production and subsequent purification of functional bacteriorhodopsin in high yield. (c) 2010 Elsevier Inc. All rights reserved.

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