基于DnaSP软件分析嗜尸性丽蝇COⅠ基因序列多态性

CO Ⅰ gene sequence polymorphisms analysis of the necrophagous calliphoridae flies based on the DnaSP software

目的 基于DnaSP软件分析嗜尸性丽蝇线粒体细胞色素c氧化酶亚基Ⅰ(Cytochrome c Oxidase SubunitⅠ,COⅠ)基因序列,以期发现潜在的小片段多态性区域,为嗜尸性蝇类尤其是降解蝇类样本的检测以及相关技术的应用提供基础数据。方法 通过NCBI数据库检索世界范围内的嗜尸性丽蝇COⅠ序列,以丽蝇科蝇类的拉丁学名和COⅠ为关键词进行索引,序列长度大于1 300 bp为限定条件。利用MEGA7.0软件比对并进行聚类,DnaSP 5.10软件分析序列多态性。结果 获得18种常见丽蝇的224条COⅠ序列,比对剪切后终序列长度为1 206bp,存在411个可变位点,滑动窗口分析结果显示,6处可变位点数目较多,分别位于185~269 bp、290~374 bp、570~689 bp、815~934 bp、955~1 039 bp、1 060~1 179 bp。结论 长度为1 206 bp的COⅠ序列中,发现6处多态性信息丰富的小片段,这为降解蝇类样本的检测以及短片段检测技术用于嗜尸性蝇类种属鉴定提供了基础资料。

Objective The mitochondrial DNA cytochrome c oxidase subunit Ⅰ(CO Ⅰ) gene sequences of the necrophagous Calliphoridae flies were analyzed based on the DnaSP software,to explore the potential polymorphic short fragments and provide basic data for the detection of degraded samples and the application of related techniques.Methods The CO Ⅰ sequences of the necrophagous Calliphoridae flies worldwide were searched in NCBI database.The Latin name of the Calliphoridae flies and CO Ⅰ were used as the key words,and the length of the sequence was refined to more than 1 300bp.The obtained sequences were aligned and analyzed by the MEGA7.0 and DnaSP 5.10 software.Results A total of 224CO Ⅰ sequences from 18 species of the Calliphoridae flies were obtained,and the final sequence length was 1 206 bp.A total of 411 variable sites were found.The results of sliding window analysis showed that there were six polymorphic regions at 185 ~ 269 bp,290 ~ 374 bp,570 ~ 689 bp,815 ~ 934 bp,955 ~ 1 039 bp,1 060 ~1 179 bp,respectively.Conclusions In this study,six polymorphic short fragments have been found in the 1 206 bp CO Ⅰ sequence,which could be applied in the degraded sample detection and in the identification of necrophagous flies using short fragments analysis.