| H3N2亚型犬流感病毒HA1基因的原核表达及抗原性分析 |
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| 引用本文： | 张敦伟,熊永忠,远立国,贾坤,张桂红,李守军.H3N2亚型犬流感病毒HA1基因的原核表达及抗原性分析[J].中国预防兽医学报,2011,33(2). |
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| 作者姓名： | 张敦伟 熊永忠 远立国 贾坤 张桂红 李守军 |
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| 作者单位： | 1. 华南农业大学兽医学院,广东,广州,510642
2. 中国农业科学院哈尔滨兽医研究所哈尔滨维科生物技术开发公司,黑龙江,哈尔滨,150069 |
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| 基金项目： | 国家自然科学基金(30972233); 广东省科技计划项目(2008A060202017、2009B060300024); 广东省自然科学基金重点项目(8251064201000008); 教育部博士点基金(20094404110017) |
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| 摘 要： | 为原核表达H3N2亚型犬流感病毒(CIV)HA1蛋白,本研究利用特异性引物扩增CIV H3分离株的HA1基因,将其克隆到pMD18-T载体后进行序列测定。再将其亚克隆于pET-32a(+)中构建重组表达质粒pET-HA1。将该质粒转化于大肠杆菌BL21(DE3)中,经IPTG诱导,SDS-PAGE电泳分析,表达的重组蛋白约为58 ku。纯化的HA1蛋白经western blot和Dot-ELISA鉴定表明,表达的重组HA1蛋白可以与H3N2亚型CIV阳性血清发生特异性反应。
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| 关 键 词： | 犬流感病毒 HA1基因 原核表达 抗原性 |
Prokaryotic expression of H3N2 canine influenza virus HA1 gene and antigenic analysis |
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| ZHANG Dun-wei,XIONG Yong-zhong,YUAN Li-guo,RA Kun,MANG Gui-hong,LI Shou-jun.Prokaryotic expression of H3N2 canine influenza virus HA1 gene and antigenic analysis[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(2). |
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| Authors: | ZHANG Dun-wei XIONG Yong-zhong YUAN Li-guo RA Kun MANG Gui-hong LI Shou-jun |
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| Institution: | ZHANG Dun-wei1,XIONG Yong-zhong2,YUAN Li-guo1,JIA Kun1,ZHANG Gui-hong1*,LI Shou-jun1*(1.College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China,2.Harbin Weike Biotechnology Development Company,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China) |
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| Abstract: | To express canine influenza virus(CIV) swbtype H3N2 HA1 protein,the HA1 gene was amplified by PCR and cloned into vector pMD18-T and then sequence determination.The HA1 gene was cloned into vector pET-32a(+).The recombinant plasmid pET-HA1 was transformed into BL21(DE3) and induced with IPTG.The expressed HA1 fusion protein had a MW of 58 ku and was purified.Western blot and Dot-ELISA showed that the purified HA1 protein could be recognized by CIV positive serum. |
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| Keywords: | canine influenza virus HA1 gene prokaryotic expression antigenicity |
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