| 甲型流感病毒H1N1亚型NS1蛋白真核表达载体的构建与表达 |
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| 引用本文： | 张文帅,卞倩,迟莹,温恬,李燕,焦永军. 甲型流感病毒H1N1亚型NS1蛋白真核表达载体的构建与表达[J]. 现代预防医学, 2011, 38(16): 3303-3305,3308 |
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| 作者姓名： | 张文帅 卞倩 迟莹 温恬 李燕 焦永军 |
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| 作者单位： | 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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| 基金项目： | 江苏省自然科学基金资助项目 (BK2009433); 江苏省卫生厅资助项目 (Z200919) |
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| 摘 要： | [目的]构建甲型流感病毒H1N1亚型NS1蛋白真核表达载体,并在293T细胞中表达.[方法]采用RT-PCR技术,从甲型流感病毒H1N1毒株提取的病毒总RNA中,扩增NS1全长基因,将其克隆至pMD18-T Vector中构建pMD18-T-NS1质粒,双酶切pMD18-T-NS1与PXJ40-HA后,构建真核表达载体PXJ40-HA-NS1,经酶切及测序鉴定后将质粒转染到293T细胞中,通过免疫印迹法鉴定NS1蛋白的表达.[结果]酶切、测序证明重组真核表达载体PXJ40-HA-NS1构建成功,免疫印迹法可见NS1基因编码蛋白表达.[结论]成功构建甲型流感病毒H1N1亚型NS1蛋白真核表达载体PXJ40-HA-NS1,并在293T细胞中传染表达,该表达载体的构建为后期建立稳定表达NS1蛋白的细胞模型和NS1蛋白功能研究提供了材料.
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| 关 键 词： | 甲型流感病毒H1N1 NS1 真核表达 免疫印迹 |
CONSTRUCTION AND EUKARYOTIC EXPRESSION OF NS1 PROTEIN OF INFLUENZA A VIRUS SUBTYPE H1N1 |
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| ZHANG Wen-shuai,BIAN Qian,CHI Ying,et al.. CONSTRUCTION AND EUKARYOTIC EXPRESSION OF NS1 PROTEIN OF INFLUENZA A VIRUS SUBTYPE H1N1[J]. Modern Preventive Medicine, 2011, 38(16): 3303-3305,3308 |
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| Authors: | ZHANG Wen-shuai BIAN Qian CHI Ying et al. |
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| Affiliation: | ZHANG Wen-shuai,BIAN Qian,CHI Ying,et al.(Institute of Pathogenic Microbiology,Jiangsu Provincial Center for Disease Prevention and Control,Nanjing 210009,China) |
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| Abstract: | [Objective]To construct the full-length NS1 gene of influenza A Virus Subtype H1N1 into a eukaryotic expression vector PXJ40-HA,and study the expression of NS1 gene in transfected 293T cells.[Methods]The NS1 gene of influenza A Virus Subtype H1N1 was amplified by RT-PCR and cloned into pMD18-T vector to construct a plasmid,named pMD18-T-NS1.The pMD18-T-NS1 and the PXJ40-HA were double digested by the same restrict enzymes,the recombinant eukaryotic expression vector PXJ40-HA-NS1 subsequently yielded. The ex... |
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| Keywords: | Influenza A Virus subtype H1N1 NS1 Eukaryotic expression Western blot |
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