| SARS冠状病毒N基因的扩增与克隆 |
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| 引用本文： | 肖维威,马文丽,张宝,王艳,毛向明,彭翼飞,宋艳斌,吴清华,郑文岭.SARS冠状病毒N基因的扩增与克隆[J].第一军医大学学报,2004,24(1):39-41. |
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| 作者姓名： | 肖维威 马文丽 张宝 王艳 毛向明 彭翼飞 宋艳斌 吴清华 郑文岭 |
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| 作者单位： | [1]第一军医大学分子生物学研究所,广东广州510515 [2]广州军区广州总医院分子肿瘤学研究所,广东广州510010 |
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| 摘 要： | 目的 RT—PCR扩增、克隆SARS冠状病毒N蛋白基因。方法 根据GenBank数据库中TOR2株的全基因组序列，利用Primer Premier5．0软件设计引物RT-PCR巢式扩增SARS冠状病毒的N基因，PCR产物克隆后进行测序鉴定。结果 序列分析表明。pMD18-T载体中已成功重组了N基因。结论 N蛋白基因的扩增、克隆成功，为N蛋白的表达、N蛋白结构与功能的研究奠定了基础。
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| 关 键 词： | SARS 严重急性呼吸综合症 传染性非典型肺炎 冠状病毒 基因扩增 基因克隆 N蛋白 |
Amplification and cloning of the N gene of SARS-associated coronavirus] |
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| Wei-wei Xiao,Wen-li Ma,Bao Zhang,Yan Wang,Xiang-ming Mao,Yi-fei Peng,Yan-bin Song,Qing-hua Wu,Wen-ling Zheng.Amplification and cloning of the N gene of SARS-associated coronavirus][J].Journal of First Military Medical University,2004,24(1):39-41. |
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| Authors: | Wei-wei Xiao Wen-li Ma Bao Zhang Yan Wang Xiang-ming Mao Yi-fei Peng Yan-bin Song Qing-hua Wu Wen-ling Zheng |
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| Institution: | Institute of Molecular Biology, First Military Medical University, Guangzhou 510515, China. |
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| Abstract: | OBJECTIVE: To amplify and clone the N gene of severe acute respiratory syndrome-associated coronavirus. METHOD: Using primer Premier 5.0 software, two pairs of nested PCR primers were designed to amplify the N gene. After purification, the amplified products were cloned into pMD18-T vectors, and the positive clones with the inserted fragments were identified by sequence analysis. RESULTS: The amplified products was about 1 375 bp in length, and sequence analysis demonstrated that the N gene fragments had been successfully inserted into pMD18-T vectors. CONCLUSION: The successful amplification and cloning of N gene facilitates further investigation of the expression of the N protein and study of its structure and functions. |
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