Wheat germ systems for cell-free protein expression
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作者:
Harbers, Matthias
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RIKEN, Ctr Life Sci Technol, Tsurumi ku, Yokohama, Kanagawa 2300045, Japan
CellFree Sci, Tsurumi ku, Yokohama, Kanagawa 2300046, JapanRIKEN, Ctr Life Sci Technol, Tsurumi ku, Yokohama, Kanagawa 2300045, Japan
Harbers, Matthias
[1
,2
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机构:
[1] RIKEN, Ctr Life Sci Technol, Tsurumi ku, Yokohama, Kanagawa 2300045, Japan
[2] CellFree Sci, Tsurumi ku, Yokohama, Kanagawa 2300046, Japan
Cell-free protein expression plays an important role in biochemical research. However, only recent developments led to new methods to rapidly synthesize preparative amounts of protein that make cell-free protein expression an attractive alternative to cell-based methods. In particular the wheat germ system provides the highest translation efficiency among eukaryotic cell-free protein expression approaches and has a very high success rate for the expression of soluble proteins of good quality. As an open in vitro method, the wheat germ system is a preferable choice for many applications in protein research including options for protein labeling and the expression of difficult-to-express proteins like membrane proteins and multiple protein complexes. Here I describe wheat germ cell-free protein expression systems and give examples how they have been used in genome-wide expression studies, preparation of labeled proteins for structural genomics and protein mass spectroscopy, automated protein synthesis, and screening of enzymatic activities. Future directions for the use of cell-free expression methods are discussed. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
机构:
Seattle Biomed Res Inst, Seattle, WA 98109 USA
Univ Washington, Dept Global Hlth, Seattle, WA 98195 USASeattle Biomed Res Inst, Seattle, WA 98109 USA
Guild, Katherine
Zhang, Yang
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Seattle Biomed Res Inst, Seattle, WA 98109 USASeattle Biomed Res Inst, Seattle, WA 98109 USA
Zhang, Yang
Stacy, Robin
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Seattle Biomed Res Inst, Seattle, WA 98109 USASeattle Biomed Res Inst, Seattle, WA 98109 USA
Stacy, Robin
Mundt, Elizabeth
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Seattle Biomed Res Inst, Seattle, WA 98109 USASeattle Biomed Res Inst, Seattle, WA 98109 USA
Mundt, Elizabeth
Benbow, Sarah
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Seattle Biomed Res Inst, Seattle, WA 98109 USASeattle Biomed Res Inst, Seattle, WA 98109 USA
Benbow, Sarah
Green, Amanda
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Seattle Biomed Res Inst, Seattle, WA 98109 USASeattle Biomed Res Inst, Seattle, WA 98109 USA
Green, Amanda
Myler, Peter J.
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Seattle Biomed Res Inst, Seattle, WA 98109 USA
Univ Washington, Dept Global Hlth, Seattle, WA 98195 USASeattle Biomed Res Inst, Seattle, WA 98109 USA