| 甲型H1N1流感病毒非结构蛋白NS1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,温恬,迟莹,李燕,焦永军. 甲型H1N1流感病毒非结构蛋白NS1真核表达载体的构建与表达[J]. 细胞与分子免疫学杂志, 2011, 27(3): 287-289 |
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| 作者姓名： | 张文帅 卞倩 温恬 迟莹 李燕 焦永军 |
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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,江苏,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H1N1流感病毒非结构蛋白NS1真核表达载体并表达其编码蛋白(转染293T细胞)。方法:从江苏首例甲型H1N1流感病毒毒株(A/Nan jing/1/2009(H1N1))提取病毒RNA,采用RT-PCR技术扩增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Western blot鉴定NS1蛋白的表达。结果:经双酶切、测序鉴定证实NS1基因的真核表达载体构建成功。West-ern blot法可见NS1基因编码蛋白的成功表达。结论:成功克隆NS1全长基因,并构建了其真核表达载体,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H1N1流感病毒 NS1 真核表达 免疫印迹 |
Construction and expression of eukaryotic expression vector of NS1 protein of influenza A(H1N1) |
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| ZHANG Wen-shuai,BIAN Qian,WEN Tian,CHI Ying,LI Yan,JIAO Yong-jun. Construction and expression of eukaryotic expression vector of NS1 protein of influenza A(H1N1)[J]. Chinese journal of cellular and molecular immunology, 2011, 27(3): 287-289 |
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| Authors: | ZHANG Wen-shuai BIAN Qian WEN Tian CHI Ying LI Yan JIAO Yong-jun |
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| Affiliation: | ZHANG Wen-shuai,BIAN Qian,WEN Tian,CHI Ying,LI Yan,JIAO Yong-jun 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: | AIM: To insert the full-length NS1 gene of influenza A(H1N1) into an eukaryotic expression vector PXJ40-HA,and to evaluate the expression of NS1 gene in transfected 293T cells.METHODS: The NS1 gene of influenza A(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 both digested using the same restrict enzymes and ligated,yielding the recombinant eukaryotic expression vector PXJ40-HA-NS1.The expression of the NS1 gene in ... |
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| Keywords: | influenza A(H1N1) NS1 eukaryotic expression Western blot |
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