Wheat Germ Systems for Cell-Free Protein Expression

FEBS Lett. 2014 Aug 25;588(17):2762-73. doi: 10.1016/j.febslet.2014.05.061. Epub 2014 Jun 12.

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.

Keywords: Automation; Cell-free; Isotopic labeling; Protein expression; Unnatural amino acid; Wheat germ.

Publication types

  • Review

MeSH terms

  • Animals
  • Cell-Free System
  • Humans
  • Protein Biosynthesis*
  • Robotics
  • Triticum / chemistry
  • Triticum / metabolism*