NR AVRX
AU Zhang,J.; Gao,J.M.; Li,F.; Han,J.; Chen,L.; Zhang,B.Y.; Wang,X.F.; Zhou,W.; Liu,Y.; Dong,X.P.
TI Establishment of cell free conversion system with biotin-labelled recombinant PrPsen expressed in E. coli.
QU Biomedical and Environmental Sciences 2006 Jun; 19(3): 214-8
PT journal article; research support, non-u.s. gov't
AB 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.
MH Animals; *Biotin; Blotting, Western; *Cell-Free System; Chromatography, Affinity/methods; Cricetinae; Escherichia coli/*genetics; Gene Expression Regulation; Molecular Weight; Peptide Hydrolases/analysis/metabolism; PrPsc Proteins/*genetics/metabolism; Prion Diseases/genetics/pathology; Recombinant Proteins/*genetics/metabolism; Staining and Labeling
AD School of Medicine, Xi'an Jiaotong University, China.
SP englisch
PO USA