丙型肝炎病毒核心蛋白基因在大肠杆菌内的表达及应用

Expression of HCV Core Protein in E. coli and Its Application

将从中国丙肝病人血清中扩增克隆的丙型肝炎病毒核心蛋白基因（４０８ｂｐ）酶切处理后插入表达载体ｐＪＬＡ５０２内，获得高表达核心蛋白的重组工程菌。将重组菌经４２℃热诱导５ｈ，ＳＤＳ－ＰＡＧＥ分析表明，表达的核心蛋白占菌体蛋白总量的２０％。经分子筛和吸附层析纯化后获得的核心蛋白，ＥＬＩＳＡ检测证实有较好的抗原性和特异性。用表达的核心抗原加用表达的ＮＳ＿３抗原（Ｃ＿３３）装配的抗－ＨＣＶ试剂盒，经用标准血清验证及与国外第二代抗－ＨＣＶ试剂盒比较，证实符合丙肝诊断试剂要求。

he cloned HCV core protein gene was inserted into the plasmid vector pJLA502 at EcoRI site,and the recombinant plasmid was transformed into E,coli DH_(5a),After inducing 5hrs at 42℃,SDS-PAGE analysis showed that the expressed core protein is twenty percent of total protein of E.coliDH5a;the purified core protein by Sephadex G200 gel filtration and Sepharose-4B-IgG affinity chro-matography has excellent antigenicity and specificity,and is good for detecting anti-HCV.