中国寄生虫学与寄生虫病杂志 ›› 2010, Vol. 28 ›› Issue (6): 17-473-475.

• 研究简报 • 上一篇    下一篇

五条蚋细胞色素C氧化酶亚基Ⅰ(COⅠ)基因的生物信息学分析

罗洪斌1,杨明2 *,陈汉彬2   

  1. 1 湖北民族学院医学院生化教研室,恩施 445000; 2 贵阳医学院生物教研室,贵阳 550004
  • 出版日期:2010-12-31 发布日期:2011-03-07

Bioinformatics Analysis of COⅠ Gene of Simulium quinquestriatum

LUO Hong-Bin1,YANG Ming2 *,CHEN Han-Bin2   

  1. 1 Department of Biochemistry,Medical School,Hubei University for Nationalities,Enshi 445000,China;
    2 Department of Biology,Guiyang Medical College,Guiyang 550004,China
  • Online:2010-12-31 Published:2011-03-07

摘要: 【提要】  以五条蚋基因组DNA为模板PCR扩增其COⅠ基因序列,将所得片段克隆入pMD18-T载体,转化大肠埃希菌DH5α,筛选阳性克隆。经PCR与双酶切鉴定,获得阳性重组质粒pMT18-T-COⅠ,测序后作序列分析和同源性比较。结果表明,从五条蚋DNA中扩增出COⅠ基因及5′端tRNA-Tyr基因和3′端tRNA-Leu基因部分片段共1 621 bp,其中COⅠ基因序列长度为1 542 bp(GenBank登录号为DQ534949),与预期相符,该基因开放阅读框编码513个氨基酸,编码蛋白等电点为5.84,相对分子质量为Mr 5 650,具有COⅠ基因的核心保守结构域。与GenBank已知基因(登录号为AY251520)的序列一致性为99%。

关键词: 五条蚋, COⅠ基因, 克隆, 生物信息学

Abstract: 【Abstract】  The genomic DNA was extracted from Simulium quinquestriatum (Sq) and its COⅠ gene was amplified by PCR. The PCR product was purified and cloned into plasmid pMD18-T vector. The recombinant plasmid was transformed into Escherichia coli DH5α and then identified by digestion with restriction enzyme and PCR amplification. The amplified frac-tions (1 621 bp) included complete COⅠ gene (1 542 bp, GenBank accession number: DQ534949), 5′ tRNA-Tyr and 3′ tRNA-Leu partial fraction. The COⅠ gene sequence had a high identity (99%) with that of S. quinquestriatum (GenBank accession number: AY251520). Bioinformatics analysis showed that the Sq-COⅠopen reading frame encoded a 513-amino acid protein with Mr 5 565, pI 5.84. Structural prediction showed this protein possessed a conservative domain of COⅠ gene.

Key words: Simulium quinquestriatum, COⅠ, Clone, Bioinformatics