| 含甲型H1N1流感病毒HA基因序列病毒样颗粒的制备 |
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| 引用本文： | 单云峰,戚宇华,单军,葛以跃,赵康辰,崔仑标,史智扬,汪华.含甲型H1N1流感病毒HA基因序列病毒样颗粒的制备[J].现代预防医学,2011,38(22):4714-4716,4719. |
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| 作者姓名： | 单云峰 戚宇华 单军 葛以跃 赵康辰 崔仑标 史智扬 汪华 |
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| 作者单位： | 卫生部肠道病原微生物重点实验室,江苏省疾病预防控制中心病原微生物研究所,南京,210009 |
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| 摘 要： | 目的]构建耐RNase酶的内含甲型H1N1流感病毒HA基因序列的病毒样颗粒。方法]构建中间载体pET32a-MS2,将甲型H1N1流感病毒的HA基因片段连接中间载体上,构建原核表达载体pET32a-MS2-HA,转化宿主菌,诱导表达制备病毒样颗粒,对病毒样颗粒进行荧光定量RT-PCR检测和稳定性实验。结果]表达载体经PCR、酶切鉴定分析后证实构建成功,荧光定量RT-PCR实验表明该病毒颗粒含有HA基因片段并且颗粒稳定性良好。结论]成功构建了含甲型H1N1流感病毒HA基因序列的病毒样颗粒且稳定性良好,有望作为甲型H1N1流感病毒RNA检测的标准品和质控品。
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| 关 键 词： | MS2噬菌体 甲型H1N1流感病毒 病毒样颗粒 原核表达 |
CONSTRUCTION OF THE VIRUS-LIKE PARTICLES CONTAINING HA GENE FRGMENTS WITH TYPE A INFLUENZ A(H1N1)VIRUS |
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| SHAN Yun-feng,QI Yu-hua,SHAN Jun,et al..CONSTRUCTION OF THE VIRUS-LIKE PARTICLES CONTAINING HA GENE FRGMENTS WITH TYPE A INFLUENZ A(H1N1)VIRUS[J].Modern Preventive Medicine,2011,38(22):4714-4716,4719. |
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| Authors: | SHAN Yun-feng QI Yu-hua SHAN Jun |
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| Affiliation: | SHAN Yun-feng,QI Yu-hua,SHAN Jun,et al.(Key Laboratory of Enteric Pathogenic Microbiology,Ministry of Health,Institute of Pathogenic Microbiology,Jiangsu Provincial Center for Disease Prevention and Control,Nanjing 210009,China) |
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| Abstract: | Objective]To construct RNase-resistant virus-like particles containing HA gene sequences of type A influenza(H1N1)virus that can be used as RNA standards and controls in RNA virus detection.Methods]The coat protein and maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32a-MS2.The hemagglutinin gene(HA)sequence of type A influenza(H1N1)virus was cloned into the downstream of pET32a-MS2 bacteriophage gene to construct ... |
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| Keywords: | Bacteriophage MS2 H1N1 influenza virus Virus-like particles Prokaryotic expression vector |
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