[1]杨　 薇,曲娟娟,于 婧,等.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,(04):90-94.[doi:10.11942/j.issn1002-2767.2009.04.0090]
　YANGWei,QU Juan-juan,YU Jing,et al.Cloning of an ORF2 Fragment of Swine HEV and Constructionof Its Plant Expression Vector[J].HEILONGJIANG AGRICULTURAL SCIENCES,2009,(04):90-94.[doi:10.11942/j.issn1002-2767.2009.04.0090]
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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

《黑龙江农业科学》[ISSN:1002-2767/CN:23-1204/S] 卷: 期数: 2009年04 页码: 90-94 栏目: 出版日期: 2009-07-10
Title:
Cloning of an ORF2 Fragment of Swine HEV and Constructionof Its Plant Expression Vector
作者:
杨　 薇 1 ; 曲娟娟 1 ; 于 婧 2 ; 李 杰 3 ; 刘　 琦4; 夏善勇 4 ; 相文华 2 ; 李柱刚 4
(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)
Author(s):
YANGWei1; 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, Hei-longjiang 150030;2.National Key Laboratory of Veterinary Biotechnology , Harbin Veterinary ResearchInstitute of Chinese Academy of Agricultural Sciences, Harbin,Heilongjiang 150001;3.Life Science Col-lege of Northeast Agricultural University ,Harbin,Heilongjiang 150030;
关键词:
戊型肝炎病毒; ORF2; 克隆; 植物表达载体
Keywords:
4.Biotechnology Research Institu-te of Heilongjiang Academy of Agricultural Sciences; Key Laboratory of Crop and Livestock MolecularBreeding of Heilongjiang ; Harbin; Heilongjiang 150086)afragment of swine HEV ; ORF2; clone; plant expression vector 。
DOI:
10.11942/j.issn1002-2767.2009.04.0090
文献标志码:
A
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。
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 ORF2gene was linked to the p35S-2300-two-T-DNA-4×Ta 1 vector on which the CaMV35S promoter was replaced by the induc-ible promoter E12.The plant expression vector p2300-ORF2-E12 was constructedand transitedinto Agrobacterium LBA440by freeze-thaw method.All the experiments have laid the foundation for the future research on transgenic plant vaccine.

备注/Memo

基金项目:黑龙江省博士后启动基金资助项目(2005037187)

第一作者简介:杨薇(1983-),女,河北省沧县人,在读硕士,主要从事分子生物学的研究。E-mail:vivi0508@sohu.com。
通讯作者:李柱刚(1972-), 男, 博士, 研究员, E-mail:lizhugang @163.com。

更新日期/Last Update: 2017-03-30