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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 | |
| 引用本文： | 杨薇,曲娟娟,于婧,李杰,刘琦,夏善勇,相文华,李柱刚.猪戊型肝炎结构基因ORF2的克隆及其植物表达载体的构建[J].黑龙江农业科学,2009(4):90-94. |
| 作者姓名： | 杨薇 曲娟娟 于婧 李杰 刘琦 夏善勇 相文华 李柱刚 |
| 作者单位： | 1. 东北农业大学资源与环境学院,黑龙江哈尔滨,150030 2. 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,黑龙江哈尔滨,150001 3. 东北农业大学生命学院,黑龙江哈尔滨,150030 4. 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,黑龙江哈尔滨,150086 |
| 基金项目： | 黑龙江省博士后启动基金 |
| 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。 |
| 关 键 词： | 戊型肝炎病毒 ORF2 克隆 植物表达载体 |
Cloning of an ORF2 Fragment of Swine HEV and Construction of Its Plant Expression Vector |
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| YANG Wei,QU Juan-juan,YU Jing,LI Jie,LIU Qi,XIA Shan-yong,XIANG Wen-hua,LI Zhu-gang.Cloning of an ORF2 Fragment of Swine HEV and Construction of Its Plant Expression Vector[J].Heilongjiang Agricultural Science,2009(4):90-94. | |
| Authors: | YANG Wei QU Juan-juan YU Jing LI Jie LIU Qi XIA Shan-yong XIANG Wen-hua LI Zhu-gang |
| Institution: | YANG Wei ,QU Juan-juan ,YU Jing ,LI Jie ,LIU Qi ,XIA Shan-yong ,XIANG Wen-hua ,LI Zhu-gang (1. Resource and Environmental Science College of Northeast Agricultural University, Harbin, Hei- longjiang 150030; 2. National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, Heilongjiang 150001;3. Life Science Col- lege of Northeast Agricultural University, Harbin, Heilongjiang 150030 ;4. Biotechnology Research Institu- te of Heilongjiang Academy of Agricultural Sciences, Key Laboratory of Crop and Livestock Molecular Breeding of Heilongjiang, Harbin, Heilongjiang 150086) |
| 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. |
| Keywords: | ORF2 afragment of swine HEV ORF2 clone plant expression vector |
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