应用mtDNA COI基因序列鉴别常见嗜尸性蝇类种属

目的探讨mtDNA基因序列对常见嗜尸性蝇类的种属鉴别应用价值。方法收集不同区域2科4属6个种30个蝇类样本,提取样本线粒体DNA后扩增COI基因序列,以琼脂糖电泳检测扩增产物并测序,以DNAMAN6.0分析软件分别截取498bp序列,用MEGA5.2软件分别进行序列分析,然后构建系统发育树,比较各地区不同种属样本的序列差异。结果 6个种属的嗜尸性蝇类30个样本mtDNA的COI基因具有一定的序列差异,种内进化分歧均数在0.1%~1.6%之间,种间进化分歧均数在2.2%~11.2%之间,6个种属通过系统发育树均可明确区分。结论 COI基因序列分析和系统发育树对嗜尸性蝇类的种属检验具有重要帮助作用,可用于现场样本的准确、快速种属鉴定。

Application of mtDNA COI genes in common sarcosaphagous flies species identification

Objective To observe the application of mtDNA COI genes in common sarcosaphagous flies species identification. Methods 30 sarcosaphagous fly samples were indentified by morphological method which collected in different regions belonging to 2 families, 4 genera and 6 species. MtDNA was extracted for the PCR amplification reaction in COI gene. The PCR products were purified through agar gel electrophoresis and sequenced. Sequences of 498 bp in COI gene were disposed by multiple-alignment software of DNAMAN. Sequences divergence of 498 bp between and within species of COI gene were processed by software of MEGA. Results It was showed that there is a certain sequence differences between the 30 samples from 6 species. The intraspecific and interspecific divergence of sequence variation ranged from 0.1% to 1.6% and 2.2% to 11.2% respectively. All the species can be identified successfully by this method. Conclusion Species identification of sarcosaphagous flies can be conducted by sequence analysis and phylogenetic tree of COI gene. This method can be effectively used in fast and accurate identification in forensic entomology.