| 重组日本乙型脑炎病毒NS1的可溶性表达及抗原性分析 |
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| 引用本文： | 周丹娜,梁望旺,郭锐,段正赢,罗明杰,杨克礼,袁芳艳,刘泽文,田永祥.重组日本乙型脑炎病毒NS1的可溶性表达及抗原性分析[J].湖北农业科学,2012(2):358-360,366. |
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| 作者姓名： | 周丹娜 梁望旺 郭锐 段正赢 罗明杰 杨克礼 袁芳艳 刘泽文 田永祥 |
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| 作者单位： | 湖北省农业科学院畜牧兽医研究所;襄阳市动物卫生监察所 |
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| 基金项目： | 湖北省动物胚胎工程及分子育种重点实验室开放课题(2011ZD152) |
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| 摘 要： | 根据日本乙型脑炎病毒(JEV)Whe株非结构蛋白质1(NS1)的基因序列,设计引物,利用反转录PCR从实验室保藏JEV Whe株中克隆NS1全长序列,并将其插入pET28a表达载体,构建重组表达质粒NS1-pET28a,转化大肠杆菌BL21(ED3),经IPTG诱导,得到可溶性表达的融合蛋白质His-NS1。结果表明,该融合蛋白质分子量为46ku,大量表达于上清中。镍离子亲和层析柱进行纯化得到His-NS1蛋白质,经Western-blot检测,证明其具有良好的免疫学活性,这为进一步研究JEV NS1的功能及应用奠定了基础。
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| 关 键 词： | 日本乙型脑炎 非结构蛋白质1 可溶性表达 抗原性分析 |
Soluble Expression and Activity Analysis of Japaness Encephalitis Virus Nonstructural Protein 1 |
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| ZHOU Dan-na,LIANG Wang-wang,GUO Rui,DUAN Zheng-ying,LUO Ming-jie,YANG Ke-li,YUAN Fang-yan,LIU Ze-wen,TIAN Yong-xiang.Soluble Expression and Activity Analysis of Japaness Encephalitis Virus Nonstructural Protein 1[J].Hubei Agricultural Sciences,2012(2):358-360,366. |
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| Authors: | ZHOU Dan-na LIANG Wang-wang GUO Rui DUAN Zheng-ying LUO Ming-jie YANG Ke-li YUAN Fang-yan LIU Ze-wen TIAN Yong-xiang |
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| Institution: | 1(1.Institute of Animal Husbandry and Veterinary,Hubei Academy of Agricultural Sciences,Wuhan 430064,China; 2.Xiangyang Animal Health Inspection Institute,Xiangyang 441000,Hubei,China) |
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| Abstract: | NS1 gene of Japanese encephalitis virus was cloned by RT-PCR and fused with His by constructing prokaryotic expression plasmid NS1-pET28a and transforming BL21(ED3).After induction by IPTG,high soluble expression of fusion protein His-NS1 was obtained and purified.SDS-PAGE analysis and Western blotting showed that the fusion protein was 46 ku and had immunological activity.The research laid the foundation for further research and investigation on function and application of JEV NS1 protein. |
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| Keywords: | Japanese encephalitis nonstructural protein 1 soluble expression antigenicity analysis |
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