Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrPsen Expressed in E. Coli
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Key words:
- Transmissible spongiform encephalopathies /
- Prion /
- Biotin /
- Cell free conversion /
- 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. Conclusion The new conversion model provides a reliable, easily handling, and environment-friendly method for studies of prion and transmissible spongiform encephalopathies.
Citation: | JIN ZHANG, JIAN-MEI GAO, FENG LI, JUN HAN, LAN CHEN, BAO-YUN ZHANG, XIAO-FAN WANG, WEI ZHOU, YONG LIU, XIAO-PING DONG. Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrPsen Expressed in E. Coli[J]. Biomedical and Environmental Sciences, 2006, 19(3): 214-218. |