| 传染性非典型肺炎病毒核蛋白的表达与活性检测 |
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| 引用本文： | 王颖彬,葛胜祥,林鉴,程通,李少伟,何志强,欧山海,张军,夏宁邵.传染性非典型肺炎病毒核蛋白的表达与活性检测[J].厦门大学学报(自然科学版),2003,42(4):416-420. |
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| 作者姓名： | 王颖彬 葛胜祥 林鉴 程通 李少伟 何志强 欧山海 张军 夏宁邵 |
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| 作者单位： | 厦门大学细胞生物学与肿瘤细胞工程教育部重点实验室,福建,厦门,361005 |
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| 基金项目： | 教育部跨世纪优秀人才培养计划资助 |
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| 摘 要： | 用PCR方法，人工合成传染性非典型肺炎病毒(SARS-CoV)核蛋白(N)全编码基因，并构建原核表达载体．在大肠杆菌中进行表达．结果重组N蛋白表达产量占菌体总蛋白的40％以上，主要以可溶形式存在．用SARS患者急性期血清进行蛋白印迹检测，表明可溶形式和包含体形式均有明显活性．包含体形式的重组蛋白经纯化后纯度可达90％以上，活性与纯化前相当，可作为SARS抗体诊断试剂盒的抗原原料．
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| 关 键 词： | 传染性非典型肺炎病毒 核蛋白 基因表达 活性检测 原核表达载体 SARS－CoV |
| 文章编号： | 0438-0479(2003)04-0416-05 |
| 修稿时间： | 2003年5月18日 |
Expression and Activity Determination of Nucleoprotein of Coronavirus Associated with Severe Acute Respiratory Syndrome |
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| WANG Yin-Bin,GE Sheng-xiang,LIN Jian,CHENG Tong,LI Shao-wei,HE Zhi-qiang,OU Shan-hai,ZHANG Jun,XIA Ning-shao.Expression and Activity Determination of Nucleoprotein of Coronavirus Associated with Severe Acute Respiratory Syndrome[J].Journal of Xiamen University(Natural Science),2003,42(4):416-420. |
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| Authors: | WANG Yin-Bin GE Sheng-xiang LIN Jian CHENG Tong LI Shao-wei HE Zhi-qiang OU Shan-hai ZHANG Jun XIA Ning-shao |
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| Abstract: | The complete coden sequence of nucleoprotein ( N ) of a novel coronavirus associated with severe acute respiratory syndrome ( SARS-CoV ) was synthesized by PCR. The N gene was cloned into a prokaryotic expression plasmid, and expressed in E. coli. The expressed recombinant N protein accounted for more than 40% quantity of bacterial total protein, and most existed as soluble protein. The Western blotting using a SARS patient acute phase serum showed that both soluble protein and insoluble protein had obvious reactivity. More than 90% purity can be obtained after purification, and the reactivity with serum remained similar to that of pre-purification, so can be used as antigen for the production of SARS antibody diagnosis kit. |
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| Keywords: | severe acute respiratory syndrome nucleoprcitein prokaryotic expression |
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