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.
引用
收藏
页码:2762 / 2773
页数:12
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