Establishment of cell free conversion system with biotin-labelled recombinant PrPsen expressed in E. coli

Biomed Environ Sci. 2006 Jun;19(3):214-8.

Abstract

Objective: To report a protocol using biotin-labelled PrP protein in cell free conversion assay instead of isotope.

Methods: A hamster PrP protein (HaPrP) was expressed in E. coli and purified with HIS-tag affinity chromatograph. After being labelled with biotin, HaPrP was mixed with PrPSc preparation from scrapie strain 263K.

Results: Protease-resistant bands were detected after four-day incubation.

Conclusion: The new conversion model provides a reliable, easily handling, and environment-friendly method for studies of prion and transmissible spongiform encephalopathies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biotin*
  • Blotting, Western
  • Cell-Free System*
  • Chromatography, Affinity / methods
  • Cricetinae
  • Escherichia coli / genetics*
  • Gene Expression Regulation
  • Molecular Weight
  • Peptide Hydrolases / analysis
  • Peptide Hydrolases / metabolism
  • PrPSc Proteins / genetics*
  • PrPSc Proteins / metabolism
  • Prion Diseases / genetics
  • Prion Diseases / pathology
  • Recombinant Proteins / genetics*
  • Recombinant Proteins / metabolism
  • Staining and Labeling

Substances

  • PrPSc Proteins
  • Recombinant Proteins
  • Biotin
  • Peptide Hydrolases