HBV S基因与HCV C基因嵌合真核表达质粒的构建及鉴定

The Constructions of Recombinant Eukaryotic Expression Plasmids Including Both HBV S Gene and HCV C Gene

目的 构建含HBVS基因及HCVC基因的嵌合基因真核表达质粒 ,为HBV和HCV嵌合基因疫苗研究奠定基础。 方法 PCR扩增分别获得HBVpreS1 +preS2 +S、preS2 +S、S基因片段及HCVC基因片段 ,以pcDNA3 .1 (+)为载体 ,构建同时含HBV preS1 +preS2 +S、preS2 +S或S基因与HCVC区基因的嵌合真核表达质粒S1S2SCpcDNA3 .1、S2SCpcDNA3 .1、SCpcDNA3 .1。 结果 经酶切鉴定及测序分析 ,所构建的嵌合表达质粒均连接正确 ,与预计一致。 结论 嵌合基因真核表达质粒的成功构建 ,将为观察嵌合基因免疫及HBV。

Objective To construct recombinant eukaryotic expression plasmids including both HBV S gene and HCV C gene, and to develop chimeric genetic vaccine of HBV and HCV. Methods Four gene fragments of HBV preS1+preS2+S, preS2+S, S and HCV C were amplified by PCR and these fragments were cloned into eukaryotic expression vector pcDNA3.1(+). Three recombinant eukaryotic expression plasmids were constructed and were named S1S2SCpcDNA3.1,S2SCpcDNA3.1,SCpcDNA3.1. These plasmids include HBV S (preS1+preS2+S, preS2+S or S) gene and HCV C gene simultaneously. Result These recombinant expression plasmids were analyzed by Enzyme digestion and sequence, three recombinant eukaryotic expression plasmids were correctly constructed. Conclusion The successful constructions of recombinant eukaryotic expression plasmids will be helpful to study chimeric genetic vaccine of HBV and HCV.