| 禽流感病毒H5N1血凝素基因和神经氨酸酶基因在真核酵母菌中的表达 |
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| 引用本文： | 邹淑梅,于在江,张烨,辛丽,陈永坤,陈禹,陈清轩,舒跃龙.禽流感病毒H5N1血凝素基因和神经氨酸酶基因在真核酵母菌中的表达[J].实验动物科学,2011,28(3):1-5. |
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| 作者姓名： | 邹淑梅 于在江 张烨 辛丽 陈永坤 陈禹 陈清轩 舒跃龙 |
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| 作者单位： | 1. 中国疾病控制中心病毒病预防控制所,北京,100052
2. 北京标凯科技有限公司,北京,100094 |
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| 基金项目： | “十一五”国家科技支撑计划(No.2006BAD06A15) |
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| 摘 要： | 目的克隆、表达和鉴定禽流感病毒H5N1血凝素基因（hemagglutinin,HA）和神经氨酸酶基因（neuramidinase,NA）序列,为制备抗体和基因工程疫苗打下基础。方法在成功克隆禽流感病毒H5N1全长HA、NA基因并测序的基础上,将部分基因序列克隆到表达载体pMET A上,构建了重组表达质粒pMET A/HA（49～1 587 bp）、pMET A/NA（121～1 200 bp）,电转化真核酵母菌pMAD16,甲醇诱导表达,利用Ni2＋亲和层析柱对重组蛋白进行纯化,并用Western Blotting和ELISA方法检测其抗原性。结果重组蛋白在酵母菌中可以高效表达,SDS-PAGE显示蛋白表达后形成了二聚体,蛋白纯度占总蛋白的95%以上,ELISA和Western Blotting实验证实,重组蛋白具有良好的抗原性。结论本研究成功克隆和表达了禽流感病毒H5N1 HA、NA基因序列,为禽流感病毒H5N1诊断试剂和疫苗的开发等进一步的研究提供了依据。
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| 关 键 词： | 禽流感病毒H5N1 血凝素 神经氨酸酶 真核酵母表达 |
Expression of the H5N1 Influenza A Virus Hemagglutinin and Neuramidinase Genes in P. methanolica |
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| ZOU Shu-mei,YU Zai-jiang,ZHANG Ye,XIN Li,CHEN Yong-kun,CHEN Yu-bao,CHEN Qin-xuan,SHU Yue-long.Expression of the H5N1 Influenza A Virus Hemagglutinin and Neuramidinase Genes in P. methanolica[J].Shiyan Dongwu Kexue,2011,28(3):1-5. |
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| Authors: | ZOU Shu-mei YU Zai-jiang ZHANG Ye XIN Li CHEN Yong-kun CHEN Yu-bao CHEN Qin-xuan SHU Yue-long |
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| Institution: | ZOU Shu-mei1,YU Zai-jiang1,ZHANG Ye1,XIN Li1,CHEN Yong-kun1,CHEN Yu-bao2,CHEN Qin-xuan2,SHU Yue-long1(1.Department of Influenza National Institute of Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 100052,China)(2.Beijing Biokit Inc.,Beijing 100094,China) |
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| Abstract: | Objective To clone,express and characterize the H5N1 avian influenza virus hemagglutinin(HA) and neuramidinase(NA) genes.Methods Expression vectors,pMET A/HA(49-1 587 bp) and pMET A/NA(121-1 200 bp) were constructed and transferred into the pMAD16 yeast.Recombinant HA and NA were expressed inducibly by methanol,and purified through Ni2+ affinity chromatography column.Antigencity of the recombinant proteins were determined using Western Blotting and ELISA.Results The recombinant genes can be highly expressed... |
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| Keywords: | H5N1 avian influenza virus Hemagglutinin Neuramidinase Yeast expression |
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