| 鸡传染性贫血病毒衣壳蛋白的原核表达及免疫原性分析 |
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| 引用本文： | 赵锋,李学花,林世武,郭文杰,杨明,徐增强,邱建东,李进,张瑞,李彦东.鸡传染性贫血病毒衣壳蛋白的原核表达及免疫原性分析[J].中国畜牧兽医,2013,40(8):34-38. |
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| 作者姓名： | 赵锋 李学花 林世武 郭文杰 杨明 徐增强 邱建东 李进 张瑞 李彦东 |
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| 作者单位： | 太原市动物卫生监督所, 山西太原 030027 |
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| 基金项目： | 太原市科技项目资助(12-0149)。 |
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| 摘 要： | 将从组织病料中提取的鸡传染性贫血病毒(chicken infectious anemia virus,CIAV)核酸进行PCR扩增,获得vp1基因,将其克隆到表达载体pET32a(+)中,构建了CIAV vp1基因重组质粒,命名为pET32a-vp1。将 pET32a-vp1转化E.coli plys,重组菌经IPTG诱导,SDS-PAGE分析结果表明,vp1基因在大肠杆菌中成功表达。纯化的蛋白质作为包被抗原,ELISA鉴定结果表明具有良好的抗原性。本试验结果为进一步研究用重组抗原制备CIAV ELISA诊断试剂盒奠定了基础。
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| 关 键 词： | 鸡传染性贫血病毒 vp1 原核表达 |
| 收稿时间： | 2012-12-11 |
Prokaryotic Expression and Immunogenicity Analysis of Chicken Infectious Anemia Virus vp1 Gene |
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| ZHAO Feng,LI Xue-hua,LIN Shi-wu,GUO Wen-jie,YANG Ming,XU Zeng-qiang,QIU Jian-dong,LI Jin,ZHANG Rui,LI Yan-dong.Prokaryotic Expression and Immunogenicity Analysis of Chicken Infectious Anemia Virus vp1 Gene[J].China Animal Husbandry & Veterinary Medicine,2013,40(8):34-38. |
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| Authors: | ZHAO Feng LI Xue-hua LIN Shi-wu GUO Wen-jie YANG Ming XU Zeng-qiang QIU Jian-dong LI Jin ZHANG Rui LI Yan-dong |
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| Institution: | Taiyuan Municipal Animal Health Supervision Institute, Taiyuan 030027, China |
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| Abstract: | We extracted chicken infectious anemia virus (CIAV) genome from liver tissue,amplified and gained vp1 gene using the polymerase chain reaction (PCR).By using the recombinant DNA technology, we digested insert DNA and pET-32a (+) with restriction enzyme BamHⅠand XhoⅠ, and then purified pET32a(+) vector DNA and insert DNA, after that ligating pET32a(+) vector DNA and insert DNA. Transform recombinant pET32a-vp1 into non-expression host E.coli DH5α competent cells. Using colony PCR to identify positive clones and verify reading frame by sequencing.Transform the plasmid pET32a-vp1 into host E.coli plys. The recombinant bacteria was induced by IPTG and analyzed with SDS-PAGE. The results showed that vp1 gene was expressed in E.coli at high level. ELISA identified target protein was vp1 fusion protein. The present study would be helpful for the development of ELISA diagnostic kit with recombinant antigen of CIAV. |
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| Keywords: | chicken infectious anemia virus(CIAV) vp1 prokaryotic expression |
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