Wheat germ cell-free expression system as a pathway to improve protein yield and solubility for the SSGCID pipeline

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Sep 1;67(Pt 9):1027-31. doi: 10.1107/S1744309111032143. Epub 2011 Aug 31.

Abstract

Recombinant expression of proteins of interest in Escherichia coli is an important tool in the determination of protein structure. However, lack of expression and insolubility remain significant challenges to the expression and crystallization of these proteins. The SSGCID program uses a wheat germ cell-free expression system as a rescue pathway for proteins that are either not expressed or insoluble when produced in E. coli. Testing indicates that the system is a valuable tool for these protein targets. Further increases in solubility were obtained by the addition of the NVoy polymer reagent to the reaction mixture. These data indicate that this eukaryotic cell-free expression system has a high success rate and that the addition of specific reagents can increase the yield of soluble protein.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell-Free System / chemistry*
  • Escherichia coli / chemistry
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Germination
  • Plant Proteins / genetics
  • Plant Proteins / isolation & purification*
  • Plant Proteins / metabolism
  • Solubility
  • Triticum / growth & development
  • Triticum / metabolism*

Substances

  • Plant Proteins