Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrP^(sen) Expressed in E.coli 被引量：1

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

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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.

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 School of Medicine State Key Laboratory for Infectious Disease Prevention and Control National Institute for Viral Disease Control and Prevention National Laboratory of Medical Molecular Biology

出处《Biomedical and Environmental Sciences》 SCIE CAS CSCD 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.

关键词 Transmissible spongiform eneephalopathies PRION BIOTIN Cell free conversion E. coli Transmissible spongiform eneephalopathies Prion Biotin Cell free conversion E. coli

分类号 R378.21 [医药卫生—病原生物学]

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