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SARS冠状病毒N基因的扩增与克隆 被引量：2

Amplification and cloning of the N gene of SARS-associated coronavirus
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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 Premier 5.0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。 Objective To amplify and clone the N g en e of severe acute respiratory syndrome-associated coronavirus. Method Using prim er Premier 5.0 software, two pairs of nested PCR primers were designed to amplif y 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 leng th, and sequence analysis demonstrated that the N gene fragments had been succe ssfully inserted into pMD18-T vectors. Conclusion The successful amplification a nd cloning of N gene facilitates further investigation of the expression of the N protein and study of its structure and functions.
作者 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
机构地区 第一军医大学分子生物学研究所 广州军区广州总医院分子肿瘤学研究所
出处 《第一军医大学学报》 CSCD 北大核心 2004年第1期39-41,共3页 Journal of First Military Medical University
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 severe acute respi ratory syndrome SARS: N-gene, coronavirus reverse transcription-polymerase cha in reaction
分类号 R346 [医药卫生—基础医学]
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参考文献5

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同被引文献14

  • 1Liu B,Cui Q,Jiang T,Ma S. A combinational feature selection and ensemble neural network method for classification of gene expression data[J]. BMC Bioinformatics,2004,5(1):136.
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  • 6Ren Q,Sato H,Murono S,et al. Epstein-Barr virus (EBV) latent membrane protein 1 induces interleukin-8 through the nuclear factor-kappa B signaling pathway in EBV-infected nasopharyngeal carcinoma cell line [J ]. Laryngoscope,2004,114(5):855-9.
  • 7Kapoor P,Frappier L. EBNA 1 partitions Epstein-Barr virus plasmids in yeast cells by attaching to human EBNA1-binding protein 2 on mitotic chromosomes [ J ]. J Virol,2003,77(12):6946-56.
  • 8Comoli P,De Palma R,Siena S,et al. Adoptive transfer ofallogeneic Epstein-Barr virus (EBV)-specific cytotoxic T cells with in vitro antitumor activity boosts LMP2-specific immune response in a patient with EBV-related nasopharyngeal carcinoma [J]. Ann Oncol,2004,15(1):113-7.
  • 9Farrell CJ,Lee JM,Shin EC,et al. Inhibition of Epstein-Barr virusinduced growth proliferation by a nuclear antigen EBNA2-TAT peptide[J]. Proc Natl Acad Sci U S A,2004,101(13):4625-30.
  • 10Macgregor PF. Gene expression in cancer:the application of microarrays[J]. Expert Rev Mol Diagn,2003,3(2):185-200.

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第一军医大学学报
2004年 第1期

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