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Efficient production and purification of functional bacteriorhodopsin with a wheat-germ cell-free system and a combination of Fos-choline and CHAPS detergents
Authors:Takahisa Genji  Yuzuru Tozawa
Institution:a Cell-Free Science and Technology Research Center and Venture Business Laboratory, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime 790-8577, Japan
b Systems and Structural Biology Center, RIKEN, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan
Abstract: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.
Keywords:TMP  transmembrane protein  CF  cell-free  FC  Fos-choline  GFP  green fluorescent protein  CHAPS  3-[(3-cholamidopropyl)dimethylammonio]propanesulfonate  DMPC  1  2-dimyristoyl-sn-glycero-3-phosphocholine  DDM  d-maltosid" target="_blank">n-dodecyl-β-d-maltosid  DHPC  1  2-dihexanoyl-sn-glycero-3-phosphocholine  PAGE  polyacrylamide gel electrophoresis
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