HCV NS2基因表达载体的构建及在E.coli中表达

HCV NS2基因表达载体的构建及在E.coli中表达 The Construction of Expression Vector of HCV NS2 and Expression in E.coli 赵月娥 ZHAO Yue-e 1 2 first-author 叶林柏 YE Lin-bai 1 2 郜金荣 GAO Jin-rong 1 2 徐进平 XU Jin-ping 1 2 刘源洁 LIU Yuan-jie 1 2 吴正辉 WU Zheng-hui 1 2 武汉大学病毒研究所 武汉大学病毒研究所 Institute of Virology,Wuhan University Institute of Virology,Wuhan University 武汉大学病毒研究所,湖北,武汉,430072 武汉大学病毒研究所,湖北,武汉,430072 应用 PCR方法从含有丙肝病毒全长非结构蛋白基因的载体 p Blue Bac2 5中扩增出全长的 N S2基因DNA片段 ,分别克隆到表达载体 p QE3 0和转座载体 p Fast Bac HTb的多克隆位点 （ MCS） .PFast NS2通过转座插入穿梭载体 Bacmid的表达盒 ;p QENS2转化 JM10 9菌株 ,诱导表达出 N端含有 6个 His的全长 NS2蛋白 ,用 Ni-NTA- agarose柱层析纯化 ,获得提纯的全长 NS2蛋白 . Full-length NS2 gene of hepatitis C virus was obtained by PCR,using plasmid pBlueBac25 harboring HCV nonstructrual gene as template.This fragment was cloned into plasmid pQE30 and transposition vector pFastBacHTb obtained pQENS2 and pFastNS2 respectively, pFastNS2 transfected DH10Bac strain and was insert into the Bacmid of Baculovirus genome by tranposition.PQENS2 was transformed JM109 strain. The NS2 protein fused to 6 His peptides at N terminus were expressed in JM109 induced by IPTG and purified by Ni-NTA-agarose. HCV NS2 构建载体 表达 纯化 HCV NS2 construction of vector expression purification Q786 2001-04-01 2021-04-01 4