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| 猪流感病毒H1N1血凝素基因和神经氨酸酶基因在大肠杆菌中的表达 | |
| 引用本文： | 张烨,于在江,唐启慧,陈禹保,辛丽,陈永坤,陈清轩,舒跃龙.猪流感病毒H1N1血凝素基因和神经氨酸酶基因在大肠杆菌中的表达[J].生物技术通报,2010(2). |
| 作者姓名： | 张烨 于在江 唐启慧 陈禹保 辛丽 陈永坤 陈清轩 舒跃龙 |
| 作者单位： | 1. 中国疾病预防控制中心病毒病所国家流感中心,北京,100052 2. 北京标凯科技有限公司,北京,100094 3. 北京中亚国瑞生物经济研究所,北京,102206 |
| 摘 要： | 克隆、表达和鉴定猪流感病毒H1N1 HA,NA基因序列,为制备抗体和基因工程疫苗打下基础。在成功克隆猪流感病毒H1N1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pET32a(+)上,全基因序列克隆到表达载体pGEX4T-1上,构建了重组表达质粒pET32a(+)/HA(截短),pET32a(+)/NA(截短),pGEX4T-1/NA,转化大肠杆菌BL21/Rosetta,IPTG诱导表达,利用Ni2+亲和层析柱和GSTrap 4B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。结果显示,重组蛋白在大肠杆菌中可以高效表达,SDS-PAGE显示其相对分子质量与预计大小一致。ELISA和Western blotting试验证实,重组蛋白具有良好的抗原性。本研究成功克隆和表达了猪流感病毒H1N1 HA、NA基因序列,为猪流感病毒H1N1诊断试剂和疫苗的开发等进一步的研究奠定了基础。 |
| 关 键 词： | 猪流感病毒H1N1 血凝素 神经氨酸酶 克隆表达 |
Expression of the Hemagglutinin and Neuramidinase Gene of Influenza A virus H1N1 in E.coli |
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| Zhang Ye,Yu Zaijiang,Tang Qihui,Chen Yubao,Xin Li,Chen Yongkun,Chen Qingxuan,Shu Yuelong.Expression of the Hemagglutinin and Neuramidinase Gene of Influenza A virus H1N1 in E.coli[J].Biotechnology Bulletin,2010(2). | |
| Authors: | Zhang Ye Yu Zaijiang Tang Qihui Chen Yubao Xin Li Chen Yongkun Chen Qingxuan Shu Yuelong |
| Institution: | Zhang Ye1 Yu Zaijiang1 Tang Qihui2 Chen Yubao3 Xin Li1 Chen Yongkun1 Chen Qingxuan2 Shu Yuelong1(1Chinese National Influenza Center,National Institute for Viral Disease Control , Prevention,Beijing 100052,2Beijing Biokit Inc.,Beijing 100094,3SinoGreen Institute for Bioeconomy,Beijing 102206) |
| Abstract: | It was to clone,express and characterize the HA and NA Protein of Swine influenza virus H1N1.On the basis of successful cloning the full length HA and NA gene,part of the gene into pET32a(+)were ligated,and full of the gene into pGEX4T-1.An expression vector pET32a(+)/HA(cut),pET32a(+)/NA(cut),pGEX4T-1/NA were constructed and expressed in E.coli BL21/rosetta induced by IPTG.Recombinant protein was purified through affinity chromatography column.Western Blotting and ELISA were used to determine the antigenic... |
| Keywords: | Swine influenza virus H1N1 Hemagglutinin Neuramidinase Clone and express |
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