Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrP^(sen) Expressed in E.coli  

作　　者：JIN ZHANG JIAN-MEI GAO FENG LI JUN HAN LAN CHEN BAO-YUN ZHANG XIAO-FAN WAMG WEI ZHOU YONG LIU XIAO-PING DONG 

机构地区：[1]School of Medicine, Xi＇an Jiaotong University, Zhuque Street 205, Xi ＇an 710061, China State Key Laboratory for Infectious Disease Prevention and Control [2]National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Yingxin Rd 100, Beijing 100052, China [3]National Laboratory of Medical Molecular Biology, Institute of Basic Medical Science, Chinese Academy of Medical Sciences and Peking Union Medical College, Dongdan Santiao 5, Beijing 100005, China

出　　处：《Biomedical and Environmental Sciences》2006年第3期214-218,共5页生物医学与环境科学（英文版）

基　　金：This work was supported by National Natural Science Foundation of China 30070038 and 30130070, National High-Tech Research and Development Program of China (863 Project) 2001AA215391, and EU Project QLRT 2000 01441.

摘　　要：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 PrP^sen 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.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 PrP^sen 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.

关 键 词：Transmissible spongiform eneephalopathies PRION BIOTIN Cell free conversion E. coli 

分 类 号：R378.21[医药卫生—病原生物学]