| 应用HA、NA、M1和M2蛋白制备H5N1高致病性禽流感病毒样颗粒 |
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| 引用本文： | 王立强,许龙,周晓巍,杨志新,陆健昇,刘冰,夏炳辉,杨津,黄培堂.应用HA、NA、M1和M2蛋白制备H5N1高致病性禽流感病毒样颗粒[J].生物技术通讯,2011,22(6):759-763. |
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| 作者姓名： | 王立强 许龙 周晓巍 杨志新 陆健昇 刘冰 夏炳辉 杨津 黄培堂 |
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| 作者单位： | 1. 军事医学科学院生物工程研究所,北京100071;沈阳军区联勤部疾病预防控制中心,辽宁沈阳110034
2. 军事医学科学院生物工程研究所,北京,100071 |
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| 摘 要： | 目的:应用重组杆状病毒表达系统制备由HA、NA、M1和M2蛋白组成的H5N1高致病性禽流感病毒样颗粒,为研究H5N1高致病性禽流感疫苗奠定基础。方法:构建能共表达A/chicken/Jilin/2003(H5N1)禽流感病毒血凝素(HA)和神经氨酸酶(NA)、A/PR/8/34(H1N1)流感病毒基质蛋白(M1)和离子通道蛋白(M2)的2个二元重组杆状病毒,共同感染HighFive细胞,同时表达HA、NA、M1和M2蛋白,使这4种蛋白在感染的细胞内自主组装成病毒样颗粒。经差速离心和蔗糖密度梯度超速离心收获病毒样颗粒,通过Western印迹鉴定病毒样颗粒的组成,透射电镜观察病毒样颗粒形态,血凝试验测定病毒样颗粒的活性。结果:HA、NA、M1、M2蛋白在昆虫细胞中共表达,并组装成病毒样颗粒;电镜观察到病毒样颗粒的形态与流感病毒一致,直径约80 nm;血凝试验显示该病毒样颗粒具有凝集鸡红细胞的活性。结论:应用该方法可以制备流感病毒样颗粒,为H5N1流感疫苗研究提供了可行方案。
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| 关 键 词： | 流感病毒 病毒样颗粒 杆状病毒 |
Highly Pathogenic H5N1 Avian Influenza Virus-Like Particles Comprised of the HA, NA, M1 and M2 Proteins |
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| WANG Li-Qiang,XU Long,ZHOU Xiao-Wei,YANG Zhi-Xin,LU Jian-sheng,LIU Bing,XIA Bing-hui,YANG Jin,HUAN Pei-tang.Highly Pathogenic H5N1 Avian Influenza Virus-Like Particles Comprised of the HA, NA, M1 and M2 Proteins[J].Letters in Biotechnology,2011,22(6):759-763. |
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| Authors: | WANG Li-Qiang XU Long ZHOU Xiao-Wei YANG Zhi-Xin LU Jian-sheng LIU Bing XIA Bing-hui YANG Jin HUAN Pei-tang |
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| Institution: | WANG Li-Qiang1,2,XU Long1,ZHOU Xiao-Wei1,YANG Zhi-Xin1,LU Jian-Sheng1,LIU Bing1,XIA Bing-Hui1,YANG Jin1,HUANG Pei-Tang1 1.Beijing Institute of Biotechnology,Beijing 100071,2.Centers of Disease Control and Prevention of Shenyang Army Area,Shenyang 110034,China |
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| Abstract: | Objective: To develop recombinant vaccine for avian influenza of the H5N1 subtype,we expressed virus-like particles(VLP) consisting of four structural proteins of influenza virus in insect cells.Methods: Influenza VLP were produced in insect cells co-infected with two recombinant baculoviruses,one expressing the hemagglutinin(HA) and the neuraminidase(NA) of A/chicken/Jilin/2003(H5N1) virus,another expressing the matrix protein(M1) and the ion-channel protein(M2) of A/PR/8/34(H1N1) virus.VLP were purified b... |
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| Keywords: | avian influenza virus virus-like particles baculovirus |
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