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Sequencing of mitochondrial DNA cytochrome oxidase subunit Ⅰ for identification of sarcosaphagous flies (Diptera) in Chengdu
Author(s): 
Pages: 101-106
Year: Issue:  1
Journal: ACTA ENTOMOLOGICA SINICA

Keyword:  食尸性苍蝇种类鉴定线粒体DNA细胞色素氧化酶亚基Ⅰ;
Abstract: 通过检测食尸性苍蝇线粒体DNA(mtDNA)上细胞色素氧化酶亚基Ⅰ(COⅠ)中278 bp基因序列,鉴定食尸性苍蝇的种类,解决依据形态学方法不能鉴定苍蝇卵的种类、很难鉴定幼虫种类的难题,作为法医鉴别食尸性苍蝇及其幼虫、卵种类依据.随机采集放置在成都地区室外草地兔尸体上的4种15个食尸性苍蝇.利用改进的小型昆虫DNA匀浆方法提取上述苍蝇mtDNA;通过Perkin-Elmer 9600扩增仪进行PCR扩增;聚丙烯酰胺非变性凝胶连续缓冲体系垂直电泳和银染显色技术进行扩增结果检测;PCR胶回收试剂盒纯化;ABI 377测序仪测序;MEGA2.1软件包进行序列分析和构建系统发育树.在双翅目食尸性苍蝇的种内进化分歧均数小于1%,种间进化分歧均数大于7%.mtDNA上COⅠ序列分析能有效地对主要的食尸性苍蝇进行种类鉴定.该检测方法快速、简便和精确,能作为法医鉴别食尸性苍蝇种类的可靠依据.
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