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猪戊型肝炎结构基因ORF2的克隆及其植物表达载体的构建

Cloning of an ORF2 Fragment of Swine HEV and Construction of Its Plant Expression Vector

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【Author】 YANG Wei1,QU Juan-juan1,YU Jing2,LI Jie3,LIU Qi4,XIA Shan-yong4,XIANG Wen-hua2,LI Zhu-gang4(1.Resource and Environmental Science College of Northeast Agricultural University,Harbin,Heilongjiang 150030;2.National Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences,Harbin,Heilongjiang 150001;3.Life Science College of Northeast Agricultural University,Harbin,Heilongjiang 150030;4.Biotechnology Research Institute of Heilongjiang Academy of Agricultural Sciences,Key Laboratory of Crop and Livestock Molecular Breeding of Heilongjiang,Harbin,Heilongjiang 150086)

【机构】东北农业大学资源与环境学院；中国农业科学院哈尔滨兽医研究所/兽医生物技术重点实验室；东北农业大学生命学院；黑龙江省农业科学院生物技术研究所/黑龙江省作物与家畜分子育种重点实验室；

【摘要】通过RT-PCR技术从一份猪粪中扩增并克隆了戊型肝炎病毒主要结构基因ORF2部分片段,大小为681 bp,将其克隆于pMD18-T载体。序列分析表明,与GenBank公布的序列一致。然后,将该基因亚克隆到植物表达载体p35S-2300-two-T-DNA-4×Ta1上,并将载体p2300上的CaMV35S启动子替换为E12启动子,构建成植物表达载体p2300-ORF2-E12。通过冻融法将重组质粒导入根癌农杆菌LBA4404中,获得了携带ORF2基因的根癌农杆菌菌株,为转基因植物工作奠定了基础。更多还原

【Abstract】 The swine hepatitis E virus(SHEV)ORF2 fragment was amplified by RT-PCR and cloned into pMD18-T vector.Sequence analysis showed that it consisted of 681 nucleotides and was identical to that of GenBank reported.Then the ORF2 gene was linked to the p35S-2300-two-T-DNA-4×Ta 1 vector on which the CaMV35S promoter was replaced by the inducible promoter E12.The plant expression vector p2300-ORF2-E12 was constructed and transited into Agrobacterium LBA440 by freeze-thaw method.All the experiments have laid the foundation for the future research on transgenic plant vaccine.更多还原

【关键词】戊型肝炎病毒； ORF2；克隆；植物表达载体；
【Key words】 afragment of swine HEV； ORF2； clone； plant expression vector；

【基金】黑龙江省博士后启动基金资助项目(2005037187)

【文献出处】黑龙江农业科学 ,Heilongjiang Agricultural Sciences , 编辑部邮箱 ,2009年04期

【分类号】S852.5
【被引频次】2
【下载频次】87

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