title: Establishment of cell free conversion system with biotin-labelled recombinant PrPsen expressed in E. coli.
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.
  
  
  CONCLUSIONS
  The new conversion model provides a reliable, easily handling, and environment-friendly method for studies of prion and transmissible spongiform encephalopathies.
authors.affiliation: School of Medicine, Xi'an Jiaotong University, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.forename: Jian-Mei
authors.initials: JM
authors.lastname: Gao
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Han
authors.forename: Lan
authors.initials: L
authors.lastname: Chen
authors.forename: Bao-Yun
authors.initials: BY
authors.lastname: Zhang
authors.forename: Xiao-Fan
authors.initials: XF
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Dong
mesh_terms: D000818:Animals; D001710:Biotin; D015153:Blotting, Western; D002474:Cell-Free System; D002846:Chromatography, Affinity / Q000379:methods; D006224:Cricetinae; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000032:analysis / Q000378:metabolism; D018620:PrPSc Proteins / Q000235:genetics* / Q000378:metabolism; D017096:Prion Diseases / Q000235:genetics / Q000473:pathology; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D013194:Staining and Labeling
chemical_list: D018620:PrPSc Proteins; D011994:Recombinant Proteins; D001710:Biotin; D010447:Peptide Hydrolases