法医学杂志

法医学杂志 ›› 2018, Vol. 34 ›› Issue (2): 114-119.DOI: 10.3969/j.issn.1004-5619.2018.02.002

• 论著 • 上一篇    下一篇

基于28S rRNA基因序列对洛阳地区嗜尸性蝇类的分子鉴定

赵琳琳1，翟仙敦1，张  振1，吕  宙2，夏志远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:2018-04-25 出版日期:2018-04-28
  • 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（11230 0410082）；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）

Molecular Identification of Sarcosaphagous Flies in Luoyang Based on 28S rRNA Gene Sequence#br#

ZHAO Lin-lin1, ZHAI Xian-dun1, ZHANG Zhen1, L?譈 Zhou2, XIA Zhi-yuan1, MO Yao-nan1   

  1. 1. School of Forensic Medicine, Henan University of Science and Technology, Luoyang 471003, China; 2. School of Criminal Investigation, Southwest University of Political Science & Law, Chongqing 400031, China
  • Online:2018-04-25 Published:2018-04-28

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摘要： 目的 通过检测嗜尸性蝇类28S rRNA基因中715 bp序列，鉴定常见嗜尸性蝇类种属，解决其形态学鉴定难题，为死亡时间推断提供技术支持。 方法 收集洛阳地区常见嗜尸性蝇类标本29只，经形态学鉴定后，用Chelex-100法提取腿部DNA，并对28S rRNA基因片段进行扩增和测序，与GenBank和EMBL数据库中的28条相应蝇种序列进行比对，用MEGA7.0软件进行序列整理，通过BLAST搜索进行序列比对，并分析所得序列碱基组成，建立种内及种间进化分歧率，构建系统发育树。 结果 形态学鉴定29只嗜尸性蝇类归属于3科5属6种。获得28S rRNA基因中715 bp的序列，在线BLAST比对结果显示相似度100%。系统发育树显示5种蝇类可以较好聚类。不同蝇种种间差异0.007~0.045，种内差异0~0.001，种间差异和种内差异没有交叉。 结论 28S rRNA靶基因序列片段对嗜尸性蝇类有良好的鉴别能力，可以作为新的嗜尸性蝇类种属鉴定遗传标记。

关键词: 法医昆虫学, 法医遗传学, 嗜尸性蝇类, 聚类分析, 28S rRNA, 死亡时间, 洛阳

Abstract: Objective To detect 715 bp sequence of 28S rRNA in sarcosaphagous flies, and to identify their common species for solving the problem of morphological identification, as well as providing technical support for postmortem interval （PMI） estimation. Methods Twenty-nine common sarcosaphagous flies were collected in Luoyang and classified by morphological characteristics. The DNA was extracted from the fly’s legs by Chelex-100 method and then the fragments of 28S rRNA were amplified and sequenced. The results were compared with twenty-eight corresponding fly species of GenBank and EMBL databases. All the sequences were analyzed by MEGA7.0 software, and sequence alignment was performed by the searching in BLAST. The nucleotide composition was analysed, and the intraspecific and interspecific genetic distance and phylogenetic tree were established. Results Twenty-nine sarcosaphagous flies were classified into 6 species of 5 genera, 3 families by morphological characteristics. In the obtained 715 bp sequence of 28S rRNA, the comparison result of online BLAST showed that the similarity was 100%. Five species were well clustered by a phylogenetic tree. Between different groups, the interspecific and intraspecific differences ranged from 0.007 to 0.045 and 0 to 0.001, respectively. Conclusion The 28S rRNA target gene sequences shows a good identification capability, which can be a new genetic marker for the identification of sarcosaphagous flies.

Key words: forensic entomology, forensic genetics, sarcosaphagous flies, cluster analysis, 28S rRNA, postmortem interval, Luoyang