Wheat-germ cell-free production of prion proteins for solid-state NMR structural studies

N Biotechnol. 2011 Apr 30;28(3):232-8. doi: 10.1016/j.nbt.2010.06.016. Epub 2010 Jul 4.


The expression of soluble, functional protein on a preparative scale poses a central challenge for structural studies. Cell-free protein expression has become a valuable alternative to cell-based methods, and allows today the expression of milligram quantities of protein. Its use is particularly attractive for NMR studies as it allows a multitude of isotopic labeling schemes. We have implemented and further developed protocols to prepare cell-free extracts from wheat germ to produce recombinant protein for solid-state NMR studies. Furthermore, we established the Renilla luciferase model to optimise and evaluate extract quality, and report first productions of the prions Ure2p and HET-s devoted to structural studies currently ongoing in our laboratories.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell-Free System / metabolism*
  • Isotope Labeling
  • Luciferases, Renilla / genetics
  • Luciferases, Renilla / metabolism
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Prions / chemistry*
  • Prions / metabolism*
  • Prions / ultrastructure
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Triticum / embryology*


  • Prions
  • Recombinant Proteins
  • Luciferases, Renilla