法医学杂志

法医学杂志 ›› 2019, Vol. 35 ›› Issue (2): 181-186.DOI: 10.12116/j.issn.1004-5619.2019.02.010

• 论著 • 上一篇    下一篇

洛阳地区9种嗜尸性丽蝇28S rRNA和COⅠ基因序列分析

赵琳琳1， 翟仙敦1， 郑哲1， 吕宙2， 李永林1， 莫耀南1   

  1. 1. 河南科技大学法医学院，河南 洛阳 471003； 2. 西南政法大学刑事侦查学院，重庆 400031
  • 发布日期:2019-04-25 出版日期:2019-04-28
  • 通讯作者: 翟仙敦，女，博士，副教授，硕士研究生导师，主要从事法医遗传学研究；E-mail：xiandunzhai@163.com
  • 作者简介:赵琳琳（1991—），女，硕士研究生，主要从事法医遗传学研究；E-mail：flyzhaolinlin@163.com
  • 基金资助:
    河南省基础与前沿技术研究计划资助项目（112300410082）；河南科技大学基金资助项目（09001309，13000914，13000696）；洛阳市法医技术鉴定重点实验室资助项目（11550002）；河南科技大学研究生创新基金资助项目（CXJJ-2016-ZR16）；河南省高等学校重点科研项目基础研究计划资助项目（18B310002）

Analysis of 28S rRNA and COⅠ Gene Sequence of Nine Necrophagous Calliphorid Flies from Luoyang#br#

ZHAO Lin-lin1, ZHAI Xian-dun1, ZHENG Zhe1, LÜ Zhou2, LI Yong-lin1, MO Yao-nan1   

  1. 1. School of Forensic Medicine, Henan University of Science and Technology, Luoyang 471003, Henan Province, China; 2. School of Criminal Investigation, Southwest University of Political Science & Law, Chongqing 400031, China
  • Online:2019-04-25 Published:2019-04-28

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摘要： 目的 评价9种嗜尸性丽蝇的细胞核28S核糖体核糖核酸（ribosomalribonucleic acid，rRNA）和线粒体细胞色素c氧化酶亚基Ⅰ（cytochrome c oxidase subunitⅠ，COⅠ）基因序列在常见嗜尸性丽蝇种属鉴定中应用的可行性，为推断死亡时间提供技术支持。 方法 收集洛阳地区常见嗜尸性丽蝇标本23只，经形态学鉴定后，提取腿部DNA，扩增并测序细胞核28S rRNA和线粒体COⅠ基因片段，通过BLAST搜索进行序列比对，并分析所得序列的碱基组成，建立种内及种间进化分歧率，用邻接法构建系统发育树。 结果 形态学鉴定23只嗜尸性丽蝇归属于5属9种。获得28S rRNA中715 bp和COⅠ基因中637 bp的序列，在线BLAST比对结果显示相似度达99%以上，系统发育树显示9种蝇类可以较好聚类，与形态学鉴定结果一致。28S rRNA基因序列的种内差异为0，种间差异为0.001~0.033；COⅠ基因序列的种内差异为0~0.008，种间差异为0.006~0.101。 结论 联合28S rRNA和COⅠ靶基因序列片段可以有效区分本研究中的9种嗜尸性丽蝇，但对于近缘种的判定需要更多遗传标记的开发和联合应用。

关键词: 法医病理学, 法医遗传学, 法医昆虫学, 嗜尸性蝇类, 丽蝇科, 28S核糖体核糖核酸, 细胞色素c氧化酶亚基Ⅰ, 死亡时间, 洛阳

Abstract: Objective To assess the feasibility of using 28S ribosomal RNA （28S rRNA） and mitochondrial cytochrome c oxidase subunit Ⅰ （COⅠ） gene sequences of nine necrophagous Calliphorid flies for the identification of common necrophagous Calliphorid flies, and to provide technical support for postmortem interval （PMI） estimation. Methods Twenty-three Calliphorid flies were collected and identified morphologically, and DNA were extracted from legs. The gene fragments of 28S rRNA and COⅠ were amplified and sequenced, then the sequence alignment was performed with BLAST. The composition of obtained sequences was analyzed and evolutionary divergence rate between species and intraspecies were established. The phylogeny tree was constructed with Neighbor-Joining method. Results The 23 necrophagous Calliphorid flies were identified to 9 species of 5 genera. The 715 bp from 28S rRNA and 637 bp from COⅠ gene were obtained and the online BLAST result showed more than 99% of similarity. The phylogeny tree showed that the necrophagous flies could cluster well into 9 groups, which was consistent with morphological identification results. The intraspecific difference in 28S rRNA was 0 and the interspecific difference was 0.001-0.033. The intraspecific difference in COⅠ was 0-0.008 and the interspecific difference was 0.006-0.101. Conclusion Combined use of 28S rRNA and COⅠ gene sequence fragments can effectively identify the nine Calliphorid flies in this study. However, for closely related blowfly species, more genetic markers should be explored and used in combination in future.

Key words: forensic pathology, forensic genetics, forensic entomology, necrophagous flies, calliphoridae, 28S ribosomalribonucleic acid （28S rRNA）, cytochrome c oxidase subunit Ⅰ （COⅠ）, postmortem interval, Luoyang