| 禽流感病毒M2蛋白的原核表达及免疫原性研究 |
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| 引用本文： | 吴仁蔚,毕丁仁.禽流感病毒M2蛋白的原核表达及免疫原性研究[J].动物医学进展,2009,30(8):9-12. |
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| 作者姓名： | 吴仁蔚 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,430076
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： | 湖北省科技攻关项目(2006AA202A05);;湖北省科技攻关应急项目(2005AA401D60) |
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| 摘 要： | 基质蛋白M2是流感病毒的3种表面抗原之一,也是流感病毒中最为保守的蛋白之一,被认为是具有交叉保护能力的流感疫苗的理想候选抗原。论文从H9N2亚型禽流感病毒感染的狗肾传代(madindarby canine kidney,MDCK)细胞中克隆全长M2基因,并采用OE-PCR(overlap extension,PCR)得到缺失跨膜区的M2基因(sM2)。将sM2基因克隆到pGEX-KG原核表达载体,在大肠埃希菌中获得表达量较高的融合蛋白GST-sM2,而且以可溶形式存在。利用亲和层析的方法纯化蛋白,经Western blot证明,GST-sM2蛋白具有良好的免疫原性。ELISA检测发现GST-sM2能与H9、H5和H7亚型禽流感病毒抗体均发生反应。进一步将纯化蛋白与佐剂混合后免疫Balb/c鼠,结果在免疫小鼠体内产生了较高滴度的特异性抗体,为开发具有交叉保护力的禽流感疫苗奠定了基础。
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| 关 键 词： | 禽流感病毒 M2蛋白 原核表达 |
Study on Prokaryotic Expression and Immunogenicity of M2 Protein of Avian influenza virus |
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| WU Ren-wei,BI Ding-ren.Study on Prokaryotic Expression and Immunogenicity of M2 Protein of Avian influenza virus[J].Progress In Veterinary Medicine,2009,30(8):9-12. |
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| Authors: | WU Ren-wei BI Ding-ren |
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| Institution: | 1.College of Food Science and Technology;Huazhong Agrilcultural University;Wuhan;Hubei;430070;China;2.College of Veterinary Medicine;Huazhong Agricultural University;China |
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| Abstract: | M2 protein,one of the three envelope surface antigens of influenza A virus,is a conserved protein and an ideal candidate antigen for cross-protective vaccines.In this study,M2 gene from MDCK cells infected with H9N2 subtype avian influenza virus was cloned and its transmembrane domain was deleted by OE-PCR(overlap extension PCR),resulting in the mutated M2 gene(sM2).The sM2 gene was subcloned into the expression vector pGEX-KG.Expression of GST-sM2 fusion protein was observed as a soluble form in E.coli.The... |
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| Keywords: | Avian influenza virus M2 protein prokaryotic expression |
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