应用16S rDNA短序列片段鉴定常见嗜尸性丽蝇  

Identification of Sarcosaphagous Calliphorid Flies by Analyzing the Sequence of 16S rDNA

作　　者：石坚[1] 郭亚东[1] 匡栩源[1] 兰玲梅[1] 蔡继峰[1,2] 王红杰[3] 

机构地区：[1]中南大学基础医学院法医学系,湖南长沙410013 [2]司法部司法鉴定科学技术研究所上海市法医学重点实验室,上海200063 [3]中山市公安局小榄分局,广东中山528415

出　　处：《法医学杂志》2012年第4期281-286,共6页Journal of Forensic Medicine

基　　金：湖南省自然科学基金资助项目(11JJ5075);上海市法医学重点实验室资助项目(KF1203)

摘　　要：目的利用16S rDNA中289bp序列对常见嗜尸性丽蝇进行种类鉴定。方法从14个省市户外采集丽蝇科3属6种共计26个蝇类样本。利用CTAB法提取蝇类胸肌DNA,对16S rDNA的289 bp基因片段进行PCR扩增,检测扩增产物,测序并上传至GenBank,按邻接法构建系统发育树,通过序列分析建立种内及种间进化分歧表。结果上述蝇类按照不同属、种分别聚类,其种内进化分歧均数均为0,种间进化分歧均数在0.3%～6.5%。结论 16S rDNA上289 bp基因序列能有效地对国内常见丽蝇进行种类鉴定,应用该片段具有快速、低耗等优点,为应用嗜尸性丽蝇推断死亡时间提供可能。Objective To explore the application of a 289 bp fragment of the 16S rDNA gene to identify various species of sarcosaphagous Calliphorid flies. Methods Twenty-six Calliphorid flies were collected from 14 Chinese provinces. All specimens were properly assigned into three genera and six species. The DNA of the pectoralis was extracted using CTAB method. Then PCR amplification was done for the 289 bp frag- ment of the 16S rDNA gene. The PCR products were then purified and sequenced, and the obtained sequences were uploaded to GenBank. The phylogenetic tree was built by the neighbor-joining method and intraspe- cific and interspecific divergences were calculated by sequence analysis. Results The above 26 sarcos- aphagous flies could be well clustered according to different genera and species. The evolutional intraspe- cific values were all zero, the evolutional interspecific variations varied from 0.3% to 6.5%. Conclusion The 289 bp fragment of the 16S rDNA of sareosaphagous flies can be effectively used to identify most of the flies at species level. This method appears to be fast and low dissipative, which might be used to es- timate postmortem interval by sarcosaphagous flies.

关 键 词：法医遗传学 昆虫学 DNA 核糖体 嗜尸性昆虫 丽蝇科 

分 类 号：DF795.2[医药卫生—法医学]