文档介绍：栏目:论著·基础研究
SARS冠状病毒N基因的扩增与克隆
作者:肖维威1;马文丽1;张宝1;王艳1;毛向明1;彭翼飞1;宋艳斌1;吴清华1;郑文岭2 (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)
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 Premier -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。
关键词:严重急性呼吸综合征;N基因,冠状病毒;逆转录-聚合酶链反应
中图分类号:Q786  文献标识码:A  文章编号:1000-2588(2004)01-0039-03
Amplification and cloning of the N gene of SARS-associated coronavirus
XIAO Wei-wei1; MA Wen-li1; ZHANG Bao1; WANG Yan1; MAO Xiang-ming1; PENG Yi-fei1; SONG Yan-bin1; WU Qing-hua1; ZHENG Wen-ling2
1Institute of Molecular Biology, First Military Medical University, Guangzhou 510515, China;
2Institute of Molecular Oncology, Guangzhou General Hospital of mand, Guangzhou 510010, China
Abstract: Objective  To amplify and clone the N gene of severe acute respiratory syndrome-associated coronavirus. Method Using primer Premier 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 essfully inserted into pMD18-T vectors. Conclusion  T