Expression of a Series of SARS-CoV nucleocapsid(N) mutants and preparation of monoclonal antibodies against N protein

Abstract

严重急性呼吸综合征(SARS)是21世纪出现的第一个急性传染病,它是由一种新型冠状病毒—严重急性呼吸综合征冠状病毒病毒(SARS-CoV)引起的。我们根据GeneBank上公布的SARS-CoV基因组序列,以Tor2为蓝本,用PCR方法,人工合成其核衣壳蛋白(N)的全编码基因。构建了融合表达的重组质粒PN422,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生物活性。重组N蛋白经金属鳌合层析纯化后纯度可达95%以上,活性与纯化前相当。设计了一系列N末端或C末端缺失或两端都缺失的...Severe acute respiratory syndrome (SARS) was the first severe infectious disease in the 21 century,which was caused by a new coronavirus ,named by WHO as SARS coronavirus (SARS-CoV). From the Genebank we got the genic sequences of Nucleocapsid (N) protein of SARS-CoV. Then according to the sequence of strain Tor2 we synthesized our genic sequence of N protein by the method of PCR. The N gen...学位:理学硕士院系专业:生命科学学院生物学系_生物化学与分子生物学学号:20012602

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