| 鸭肝炎病毒VP1和VP3基因的克隆及其杆状病毒表达载体的构建 |
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| 引用本文： | 魏春娅,李日顺,黄昌力,张桂红.鸭肝炎病毒VP1和VP3基因的克隆及其杆状病毒表达载体的构建[J].中国兽医科学,2012(2):171-175. |
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| 作者姓名： | 魏春娅 李日顺 黄昌力 张桂红 |
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| 作者单位： | 华南农业大学兽医学院;河南科技大学 |
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| 基金项目： | 国家自然科学基金项目(30871877) |
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| 摘 要： | 为了构建鸭肝炎病毒VP1和VP3结构蛋白基因的杆状病毒表达载体,以RT-PCR方法扩增鸭肝炎病毒的VP1和VP3基因,克隆至pGEM-T载体。经筛选测序验证后,以BamHⅠ+XhoⅠ双酶切回收VP1和VP3目的片段,与经相同方法处理的杆状病毒转座载体pFastBac1连接,得到重组质粒pFastBac1-VP1和pFastBac1-VP3。将经酶切及测序鉴定的阳性重组质粒转化入含穿梭载体Bacmid的感受态细胞DH10Bac,发生转座作用,经抗性及蓝白斑筛选,成功获得各自携带VP1和VP3基因的重组穿梭载体,将其命名为rBacmid-VP1、rBacmid-VP3。研究结果为进一步在昆虫细胞中表达VP1或VP3基因,开发研制鸭肝炎病毒基因工程疫苗奠定了基础。
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| 关 键 词： | 鸭肝炎病毒 克隆 杆状病毒表达系统 载体构建 |
Cloning of VP1 and VP3 genes of DHV and construction of their baculovirus expression vectors |
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| WEI Chun-ya,LI Ri-shun,HUANG Chang-li,ZHANG Gui-hong.Cloning of VP1 and VP3 genes of DHV and construction of their baculovirus expression vectors[J].Veterinary Science in China,2012(2):171-175. |
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| Authors: | WEI Chun-ya LI Ri-shun HUANG Chang-li ZHANG Gui-hong |
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| Institution: | 1 (1.College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China; 2.Henan University of Science and Technology,Luoyang 471000,China) |
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| Abstract: | VP1 and VP3 genes of duck hepatitis virus were amplified by RT-PCR and cloned into the pGEM-T vector. After certified by sequencing and being digested with BamHⅠand XhoⅠ respectively,the genes were linked to the baculovirus transfer vector pFastBac1. In result,the recombinant vectors pFastBac1-VP1 and pFastBac1-VP3 were constructed successfully.Then the recombinant vectors were transformed into DH10Bac E.coli,with the positive recombinant bacmid rBacmid-VP1 and rBacmid-VP3 screened according to the resistant and the blue-white plague screening. The study provided theoretical and practical foundations for the expression of VP1 or VP3 genes in insect cells. |
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| Keywords: | duck hepatitis virus cloning baculovirus expression vector system vector construction |
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