【摘要】 【目的】构建含乙型肝炎病毒 (HBV)编码核心蛋白 (HBcAg)的核心 (C)基因的真核表达重组体 ,以进行核酸免疫及相关研究。【方法】以含HBV(ayw亚型 )全基因序列的质粒pHBV1为模板 ,用PCR的方法获得编码HBcAg的C基因片段(nt1889～ 2 45 7) ,该基因片段两端设计了BamHⅠ和EcoRⅠ的限制性内切酶位点 ,以便通过定向克隆将其插入真核表达质粒载体pcDNA3相应的多克隆位点。【结果】经BamHⅠ和EcoRⅠ酶切后琼脂糖凝胶电泳鉴定 ,以及重组体插入片段的DNA序列测定 ,证实成功地构建了含C基因的真核表达重组体pcDNA3 HBc。【结论】pcDNA3 HBc的构建成功为研究其表达及进行动物免疫研究提供了物质基础。更多还原

【Abstract】 Objective To creat Hepatitis B virus C gene eukaryotic expression construct, in order to study DNA mediated immunization in mouse and related issues research. Methods Using plasmid pHBV1 which contained hepatitis B virus (ayw subtype) whole DNA sequences as the source of viral genes, HBV DNA fragment(nt1889～2457) encoding for nucleocapsid protein (HBcAg) was polymerase chain reaction cloned as BamHⅠ EcoRⅠfragment. The gene was inserted into the multiple cloning site of eukaryotic expression plasmid vector pcDNA3. Result The recombinant plasmid was cleaved with EcoRⅠ and BamHⅠ. Agarose gel electrophoresis,and sequencing the insert fragment in pcDNA3 showed that pcDNA3 HBVc eukaryotic expression construct is successfully created.. Conclusion The recombinant construct pcDNA3 HBc provides available conditions for further study about it’s expression and immune response.更多还原