中文摘要：

目的制备表达诺如病毒衣壳蛋白的重组人3型腺病毒。方法将诺如病毒衣壳蛋白基因（Noro-orf2）克隆到腺病毒穿梭载体pBSE3CMV-esfp上，与线性化人3型腺病毒骨架质粒pBRAdv3共电转化感受态大肠杆菌BJ5183，使其在细菌内发生同源重组，带Noro—orf2基因的表达框置换腺病毒E3区，PCR及酶切筛选得到重组腺病毒质粒，将重组腺病毒质粒转染Hep-2细胞进行包装，获得感染性的重组腺病毒粒子，免疫组化分析重组腺病毒中诺如病毒衣壳蛋白的表达。结果同源重组后经酶切和PCR鉴定证明插入Noro—orf2基因的重组腺病毒质粒pBRAdv3E3dNor成功构建，并经转染包装得到高滴度的重组腺病毒Adv3E3dNor，免疫组化证明诺如病毒衣壳蛋白得到表达。结论成功构建表达诺如病毒衣壳蛋白的重组3型腺病毒Adv3E3dNor，为研制人3型腺病毒-诺如病毒双价疫苗奠定了基础。

英文摘要：

Objective To prepare recombinant human adenovirus type 3 expressing Norovirus capsid protein gene（Noro- orf2）. Methods The cDNA for Noro-orf2 was amplifed by RT-PCR from stool of infantile gastroenteritis and cloned into the adenovirus shuttle vector pBSE3CMV-egfp. The vector pBSE3CMV- Nor was linearized with EcoRV and Not Ⅰ , and transformed into E. coil BJ5183 with lined adenovirus genomic DNA plasmid pBRAdv3 by Rsr Ⅱ. The identification of recombinant adenovirus plasmid pBRAdv3E3dNor was performed by PCR, enzyme digestion and DNA sequencing. Then pBRAdv3E3dNor was digested with AsiS Ⅰ and transfected into Hep-2 cells with LipofectAMINETM 2000 to package recombinant adenovirus particles. Results Noro-orr2 was successfully inserted into the shuttle vector. The recombinant adenoviral plasmid pBRAdv3E3dNor was generated by homologous recombination in E. coil BJ5183 and confirmed by PCR and enzyme digestion. The recombinant adenovirus was successfully packaged and purified. Norovirus capsid protein gene expression was confirmed in Hep-2 cells by immunocytochemistry assay. Conclusion The recombinant type 3 adenovirus expressing Norovirus capsid protein gene was successfully constructed. This study laid a foundation for developing vaccine against Norovirus.