title: Wheat germ systems for cell-free protein expression.
abstract: 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.
authors.affiliation: RIKEN Center for Life Science Technologies, Suehiro-cho, Tsurumi-ku, Yokohama City, Kanagawa 230-0045, Japan; CellFree Sciences Co., Ltd., 75-1, Ono-cho, Leading Venture Plaza 201, Tsurumi-ku, Yokohama, Kanagawa 230-0046, Japan. Electronic address: matthias.harbers@riken.jp.
authors.forename: Matthias
authors.initials: M
authors.lastname: Harbers
mesh_terms: D000818:Animals; D002474:Cell-Free System; D006801:Humans; D014176:Protein Biosynthesis; D012371:Robotics; D014908:Triticum / Q000737:chemistry / Q000378:metabolism*
keywords: Automation; Cell-free; Isotopic labeling; Protein expression; Unnatural amino acid; Wheat germ