丙型肝炎病毒基因组C区、NS3区、NS4区基因的融合、表达及初步应用  

Gene Chimeric Fusion and Expression of Nucleocapsid,NS3 Regions and NS4 Regions of Hepatitis C Virus Genome

作　　者：沈小川[1] 李铭[2] 吴祥甫[1] 

机构地区：[1]中国科学院上海生物化学研究所,上海200031 [2]中国农业科学院上海家畜寄生虫病研究所,农业部动物寄生虫重点开放实验室,上海200232

出　　处：《生物工程学报》2001年第1期46-49,共4页Chinese Journal of Biotechnology

摘　　要：通过PCR技术从HCV全长基因组中扩增并克隆到Core区、NS4区的部分优势表面抗原肽的基因和C33a基因。再将它们以Core、CC3c、NS4的顺序和C33c、NS4的顺序分别拼接融合成融合抗原CCN、CN。片段之间以 Gly Gly Gly Gly 柔性连接肽连接。经核苷酸序列分析表明 ,拼接点序列正确。将融合抗原CN、CCN基因分别克隆至T7启动子控制下的表达质粒PET 2 4(a) +、PET 2 2 (b) +中 ,转化至大肠杆菌BL2 1(DE3)后 ,经IPTG诱导可高表达分子量约为 43kD、5 8kD的融合抗原 ,表达产物以包涵体形式存在。表达产物经Ni2 + IDA亲和柱纯化后 ,并对融合抗原CN、CCN的抗原性做了初步的鉴定。Genes encoding HCV core and NS4 antigen epitopes and C33c antigen were cloned from HCV genome by PCR,respectively.Two fused genes were constructed.One contained these three genes,another contained genes encoding C33c antigen and NS4 antigen epitopes.These fused genes were cloned into expression plasmid pET\|24(a)+ and pET\|22(b)+ under T7 promoter and transformed into \%E.coli\% BL21 (DE3) respectively.SDS\|PAGE analysis revealed that these fused antigens CCN\,CN were highly expressed after the induction by 1mmol/L IPTG.These Expression products were detected by western blotting with anti\|HCV serum.

关 键 词：丙型肝炎病毒 融合蛋白 基因组 表达 

分 类 号：Q78[生物学—分子生物学]