丙型肝炎病毒核心区截短型基因真核表达载体的构建及其在中国仓鼠卵巢细胞中的表达

Construction of eukaryotic expressing vector of the truncated gene of hepatitis C virus Core and expression in chinese hamater ovarg cell

目的:建立截短的丙型肝炎病毒(HCV)核心蛋白的真核表达载体,并在中国仓鼠卵巢(CHO)细胞中瞬时表达。方法:应用多聚酶链反应(PCR),以HCV-H株全长cDNA序列为模板,扩增获得C区羧基端部分缺失的基因片段,克隆入真核表达载体pcDNA3.1(-),并通过脂质体转染至CHO细胞。结果:构建了真核表达载体pcDNA3.1-C t,成功转染CHO细胞,间接免疫荧光和RT-PCR证实pcDNA3.1-C t在CHO细胞中表达截短型的C蛋白。结论:成功构建羧基端部分缺失的HCV C区基因的真核表达载体,瞬时转染CHO细胞,为进一步基因免疫和构建多表位卡介苗(BCG)活载体疫苗奠定基础。

Objective：To construct a eukaryotic expressing vector of HCV truncated Core gene and expression in chinese hamater ovarg（CHO） cell Methods：The cDNA sequence encoding truncated HCV Core gene which lack parts of the carboxyl-terminal were amplified by PCR and inserted into plasmid pcDNA3.1 （-） , transfecting CHO with liposome. Results：Construct the eukaryotic expression vector pcDNA3.1-Ct, succeeding to transfect CHO and transiently express truncated HCV core proten, which was assessed by IFA and RT-PCR Conclusion：Constructing a eukaryotic expression vector of HCV truncated Core gene, transiently transfected CHO cell and expressed truncated HCV core proten.