摘要
应用重组DNA技术构建了牦牛和双峰驼重组朊蛋白的4个表达载体,即pGEX-BoPrP(23~242)、pGEX—BoPrP(100~242)、pGEX—CaPrP(25~241)和pGEX—CaPrP(93~241),经转化E.coli BL21(DE3),成功地获得了重组菌E.coli BoPrP(23~242)、E.coli BoPrP(100~242)、E.coli CaPrP(25~241)和E.coli CaPrP(93~241),分别可表达大小约为50.2、41.7、49.9和42.4ku的融合蛋白。各重组菌经1.0mmol/L的IPTG于37℃诱导5~6h可产生占细菌总蛋白量30%~45%的融合蛋白。免疫印迹分析表明,除融合蛋白GST—BoPrP(100~242)外,GST—BoPrP(23~242)、GST—CaPrP(25~241)和GST—CaPrP(93~241)均可与抗牛单克隆抗体4C11反应,显示中国黄牛PrP^c与牦牛和双峰驼PrP^c具有共同的抗原成分。
By using the recombinant DNA technology,four expression plasmids,pGEX-BoPrP(23-242),pGEX-BoPrP(100-242),pGEX-CaPrP(25-241)and pGEX-CaPrP(93-241),of yak and camel recombinant prion proteins were constructed,After transforming E.coli BL21(DE3),the recombinant bacteria E.coli BoPrP(23-242),E.coli BoPrP(100-242),E.coli CaPrP(25-241)and E.coli CaPrP(93-241)were obtained.While the recombinant bacteria were induced by 1.0 mmol/L by IPTG at 37°C for 5 h to 6 h,the expectant fusion proteins with molecular sizes of approximately 50.2 ku,41.7 ku,49.9 ku and 42.4 ku were produced,respectively,and the expression products accounted for 30%-45%of overall bacterium protein.Western-blotting analysis confirmed that the fusion proteins,GST-BoPrP(23-242),GST-CaPrP(25-241)and GST-CaPrP(93-241)could react with the anti-bovine PrP monoclonal antibody 4Cll,with the exception of GST BoPrP(100-242).Results showed that the yak PrP and the camel PrP shared the same antigen component as Chinese yellow cattle PrP^c.
作者
吴润
陈豪泰
刘湘涛
谢庆阁
WU Run;CHEN Hao-tai;LIU Xiang-tao;XIE Qing-ge(Key Laboratory of Animal Virology,Ministry of Agriculture/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;College of Veterinary Medicine,GansuAgricultural University,Lanzhou 730070,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2006年第1期1-6,共6页
Chinese Veterinary Science
基金
农业部畜禽病毒学重点实验室基金项目(2002-01)
兰州兽医研究所所长基金项目(2004-2005)
关键词
朊粒
牦牛
双峰驼
原核细胞表达
prion
domestic yak
Camelus bactrianus
prokaryotic expression
