Establishment of cell free conversion system with biotin-labelled recombinant PrPsen expressed in E. coli.
- Author:
Jin ZHANG
1
;
Jian-Mei GAO
;
Feng LI
;
Jun HAN
;
Lan CHEN
;
Bao-Yun ZHANG
;
Xiao-Fan WANG
;
Wei ZHOU
;
Yong LIU
;
Xiao-Ping DONG
Author Information
- Publication Type:Journal Article
- MeSH: 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
- From: Biomedical and Environmental Sciences 2006;19(3):214-218
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo report a protocol using biotin-labelled PrP protein in cell free conversion assay instead of isotope.
METHODSA 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.
RESULTSProtease-resistant bands were detected after four-day incubation.
CONCLUSIONThe new conversion model provides a reliable, easily handling, and environment-friendly method for studies of prion and transmissible spongiform encephalopathies.
