| 禽流感病毒HA1蛋白的原核表达及其免疫原性的初步研究 |
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| 引用本文： | 吴仁蔚,张万坡,胡思顺,肖运财,毕丁仁.禽流感病毒HA1蛋白的原核表达及其免疫原性的初步研究[J].动物医学进展,2007,28(8):5-9. |
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| 作者姓名： | 吴仁蔚 张万坡 胡思顺 肖运财 毕丁仁 |
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| 作者单位： | 1. 华中农业大学食品科技学院,湖北武汉,430070
2. 华中农业大学动物医学院,湖北武汉,430070 |
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| 基金项目： | 湖北省科技攻关重大专项基金
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湖北省科技攻关应急专项基金 |
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| 摘 要： | 从H9N2亚型禽流感病毒感染的鸡胚尿囊液中提取病毒RNA,RT-PCR克隆全长血凝素(Hemagglutinin,HA)基因,并进一步克隆HA的主要抗原区HA1基因.将HA1克隆到原核表达载体pGEXKG,与谷胱苷肽(GST)融合表达,表达的融合蛋白GST-HA1以包涵体形式存在.包涵体经变性、复性处理,Western blot分析表明,表达蛋白具有良好的免疫学活性.ELISA检测发现,GST-HA1只能与H9亚型禽流感病毒抗体发生反应,而与H5和H7亚型禽流感病毒抗体无交叉反应.进一步将纯化的融合蛋白与佐剂混合后免疫Balb/c小鼠,免疫小鼠体内产生了较高滴度的特异性抗体.制备的HA1蛋白特异性强,具有良好的免疫原性,为禽流感病毒的鉴别诊断和禽流感疫苗开发奠定了基础.
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| 关 键 词： | 禽流感病毒 GST-HA1融合蛋白 原核表达 |
| 文章编号： | 1007-5038（2007）08-0005-05 |
| 修稿时间： | 2007-05-16 |
Prokaryotic Expression and Immunogenicity of HA1 Protein of Avian influenza virus |
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| WU Ren-wei,ZHANG Wan-po,HU Si-shun,XIAO Yun-cai,BI Ding-ren.Prokaryotic Expression and Immunogenicity of HA1 Protein of Avian influenza virus[J].Progress In Veterinary Medicine,2007,28(8):5-9. |
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| Authors: | WU Ren-wei ZHANG Wan-po HU Si-shun XIAO Yun-cai BI Ding-ren |
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| Institution: | 1.College of Veterinary Medicine,Huazhong Agrilcultural University,Wuhan,Hubei,430070,China;2.College of Food Science and Technology,Huazhong Agricultural University,Wuhan,Hubei,430070,China |
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| Abstract: | The HA gene was cloned by RT-PCR from allantoic fluid infected with H9N2 AIV.The major antigen domain HA1 was cloned from the full-length HA and further subcloned to the prokaryotic expression vector pGEX-KG resulting in the recombinant plasmid pKG-HA1 which was used to express the fusion protein GST-HA1 in E.coli.The immunocompetence of the purified GST-HA1 was demonstrated by Western blot and ELISA.The results suggested that the fusion GST-HA1 proteins could only react with the positive antibodies against H9 subtypes AIV but not with H5 and H7 antibodies.The high titer antibodies from Balb/c mice vaccinated with the commixture of the purified expression products and adjuvant could specifically react with H9N2 AIV.The HA1 protein in this study layed a foundation for the distinguishable diagnosis and the vaccine's development. |
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| Keywords: | Avian influenza virus GST-HA1 protein prokaryotic expression |
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