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

Bioinformatics Analysis of COⅠ Gene of Simulium quinquestriatum

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M_r5 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%。

The genomic DNA was extracted from Simulium quinquestriatum （Sq）and its COⅠgene was amplified by PCR.The PCR product was purified and cloned into plasmid pMD18-T vector.The recombinant plasmid was transformed into Escherichia coli DH5αand then identified by digestion with restriction enzyme and PCR amplification.The amplified fractions （1 621 bp）included complete COⅠgene （1 542 bp,GenBank accession number：DQ534949）,5＇ tRNA-Tyr and 3＇ tRNA-Leu partial fraction.The COⅠgene 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 M_r 5 565,pI5.84.Structural prediction showed this protein possessed a conservative domain of COⅠgene.