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Wheat germ systems for cell-free protein expression
被引:84
|作者:
Harbers, Matthias
[1
,2
]
机构:
[1] RIKEN, Ctr Life Sci Technol, Tsurumi ku, Yokohama, Kanagawa 2300045, Japan
[2] CellFree Sci, Tsurumi ku, Yokohama, Kanagawa 2300046, Japan
关键词:
Wheat germ;
Cell-free;
Protein expression;
Automation;
Isotopic labeling;
Unnatural amino acid;
HIGH-THROUGHPUT CLONING;
FREE TRANSLATION;
AMINO-ACIDS;
FUNCTIONAL-ANALYSIS;
MASS-SPECTROMETRY;
HIGHLY EFFICIENT;
MESSENGER-RNA;
PURIFICATION;
LIBRARY;
ROBUST;
D O I:
10.1016/j.febslet.2014.05.061
中图分类号:
Q5 [生物化学];
Q7 [分子生物学];
学科分类号:
071010 ;
081704 ;
摘要:
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.
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页码:2762 / 2773
页数:12
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