| 西尼罗病毒NS1蛋白的原核表达及其免疫原性研究 |
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| 引用本文： | 王彦芳,杨鹏飞,张丽萍,平芮巾,刑福彬,咸庆杰,胡孔新.西尼罗病毒NS1蛋白的原核表达及其免疫原性研究[J].检验检疫科学,2011(3):34-37,15. |
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| 作者姓名： | 王彦芳 杨鹏飞 张丽萍 平芮巾 刑福彬 咸庆杰 胡孔新 |
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| 作者单位： | 德州市中心血站;中国检验检疫科学研究院; |
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| 基金项目： | 国家公益性行业科研专项(200810162); 国家质检总局科技项目(2008IK210); 传染病国家科技重大专项(2011ZX10004-001) |
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| 摘 要： | 目的]采用大肠杆菌表达系统表达西尼罗病毒NS1蛋白,并对表达产物进行免疫原性分析。方法]将WNV NS1基因克隆入pET30a载体,获得重组表达质粒pET30a-WNV-NS1,并转化入BL21感受态细胞,将表达产物进行SDS-PAGE和W estern-blot分析,经N i＋亲和层析纯化、阴离子交换层析后获得纯化的WNV NS1蛋白;将纯化产物免疫新西兰兔,获得的免疫血清作ELISA检测。将重组蛋白包板,检测FOCUS公司商品化试剂盒中的阳性血清。结果]（1）重组WNV NS1蛋白获得表达并纯化成功。（2）Western blot结果显示表达的蛋白是带有H is标签的目的蛋白。（3）兔免疫血清的间接ELISA结果显示血清效价达到1：50000以上。（4）检测商品化试剂盒中的阳性血清,其OD450吸光值与阴性血清吸光值之比大于2。结论]重组NS1蛋白能在大肠杆菌系统中获得表达;纯化产物具有免疫原性,而且可以作为抗原检测西尼罗病毒感染的血清,为进一步研究NS1蛋白的生物学特性及西尼罗病毒的诊断试剂奠定了基础。
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| 关 键 词： | 西尼罗病毒 非结构蛋白NS1 原核表达 免疫原性 |
Expression of Protein NS1 of West Nile Virus and Identification of Its Immunogenicity |
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| Wang Yanfang,Yang Pengfei,Zhang Liping,Ping Ruijin,Xing Fubin,Xian Qingjie,Hu KongXin.Expression of Protein NS1 of West Nile Virus and Identification of Its Immunogenicity[J].Inspection and Quarantine Science,2011(3):34-37,15. |
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| Authors: | Wang Yanfang Yang Pengfei Zhang Liping Ping Ruijin Xing Fubin Xian Qingjie Hu KongXin |
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| Institution: | Wang Yanfang1,Yang Pengfei2,Zhang Liping2,Ping Ruijin2,Xing Fubin2,Xian Qingjie2,Hu KongXin2(1.Dezhou Blood Center,Dezhou,Shandong,253000,2.Chinese Academy of Inspection and Quarantine) |
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| Abstract: | To express protein NS1 of West Nile virus with pET30a and identify immunogenicity of the products,WNV NS1 gene was cloned into the pET30a vector and then the recombinant plasmid pET30a-WNV-NS1 transformed into E.coli BL21.The expressed products were analyzed with SDS-PAGE and Western blot.Purified protein WNV NS1 was obtained by Ni-NTA chromatography and Ion-exchange chromatography.Rabbits were immunized with the purified protein and the immue serum was assayed with ELISA.12 eight-well polystyrene microwell... |
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| Keywords: | West Nile Virus Nonstructural Protein NS1 Prokaryotic Expression Immunogenicity |
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