丙型肝炎病毒C区和NS3区嵌合基因的克隆及表达  

Cloning and Expression of HCV C-NS3 Chimeric Gene

作　　者：彭祥兵[1] 余健[1] 黄仕和[1] 周志军[1] 李健[1] 朱华松[1] 张爱华[1] 闭兰[1] 赵亚杰[1] 余模松[1] 

机构地区：[1]武汉生物制品研究所,武汉430060

出　　处：《中国生物制品学杂志》2006年第5期457-460,共4页Chinese Journal of Biologicals

摘　　要：目的获得丙型肝炎病毒(HCV)C-NS3嵌合抗原,以提高HCV酶联免疫吸附试验诊断试剂的质量。方法应用SOE-PCR(拼接重叠延伸PCR)的方法,构建了HCV的C区和NS3区的嵌合基因,克隆于表达载体pGEX-4T3,并转化于大肠杆菌BL21,经筛选,IPTG诱导HCV的C-NS3嵌合抗原的表达。经SDS-PAGE检测表达水平,Western blot检测抗原特异性。结果SOE-PCR构建的HCV C区和NS3区嵌合基因在BL21中获得表达。经鉴定,其相对分子质量约为75000,并具有高度特异性。结论已获得HCV C-NS3嵌合抗原,为提高HCV酶联免疫吸附试验诊断试剂的质量奠定了基础。Objective To obtain the C-NS3 antigen of HCV and improve the quality of ELISA diagnostic kit for HCV. Methods Construct the C-NS3 chimeric gene of HCV by SOE-PCR,clone into expression vector pGEX-4T3 and transform to E. coli BL21 for expression under induction of IPTG. Determine the expression level by SDS-PAGE and the specificity of expressed protein by Western blot. Results HCV C-NS3 chimeric antigen, with a relative molecular weight of 75 000, was successfully expressed and showed good specificity. Conclusion HCV C-NS3 chimeric antigen was obtained, which laid a foundation of improving the quality of ELISA diagnostic kit for HCV.

关 键 词：拼接重叠延伸PCR 嵌合基因 丙型肝炎病毒 

分 类 号：R512.63[医药卫生—内科学]