Wheat germ systems for cell-free protein expression

作者：Harbers Matthias^*

来源：FEBS LETTERS, 2014, 588(17): 2762-2773.

DOI：10.1016/j.febslet.2014.05.061

• Wheat germ
• Cell-free
• Protein expression
• Automation
• Isotopic labeling
• Unnatural amino acid

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.

• 出版日期2014-8-25