›› 2017, Vol. 35 ›› Issue (02): 273-279.

• • 上一篇    

12S rRNA、Cyt b基因标记在几类野生动物检材鉴定中的差异

黄娅琳   

  1. 南京森林警察学院鉴定中心
  • 收稿日期:2016-12-02 修回日期:2017-05-14 出版日期:2017-06-30 发布日期:2018-11-29
  • 通讯作者: 黄娅琳 E-mail:huangyalin228@163.com
  • 基金资助:
    野生动物复合基因分子标记的开发及其在近缘物种鉴定中的应用

The Differences of 12S rRNA and Cyt b Gene Marker in Wildlife Species Identification

  • Received:2016-12-02 Revised:2017-05-14 Online:2017-06-30 Published:2018-11-29

摘要: 目的 比较线粒体DNA上的12S rRNA和Cyt b基因分子标记在野生动物物种鉴定中的效果差异。方法 提取各类野生动物样本DNA,PCR扩增线粒体DNA上的12S rRNA基因片段和Cyt b基因片段,对PCR产物进行电泳检测及测序分析,测序结果在GenBank上进行BLAST搜索,同源性分析。并将实验序列和NCBI上下载的与该序列存在同源性的序列用软件MEGA 7.0进行比对并构建进化树。结果 从各类样本中成功地提取到了基因组总DNA,并成功扩增出了用于动物种属鉴定的12S rRNA基因片段、Cyt b基因片段。测序分析结果表明用12S rRNA基因片段和Cyt b基因片段均可准确鉴定黑熊、藏羚羊、鬣羚、穿山甲样本的动物种属,但12S rRNA基因片段无法准确鉴定北山羊样本,BLAST搜索结果为其近缘物种山羊,而Cyt b基因片段却可准确鉴定北山羊。各样本基于12S rRNA、Cyt b基因序列构建进化树的结果与BLAST搜索结果相符。结论 单一位点基因标记对某些近缘物种的鉴别能力较差,野生动物物种鉴定宜选用2个以上的基因标记位点。

关键词: 野生动物, 种属鉴定, 12S rRNA, Cyt b, 基因标记

Abstract: Abstract: Objective To Identify samples of different wildlife species, and compare the difference of mtDNA 12S rRNA and mtDNA Cyt b gene markers in wildlife species identification. Method The isolation of DNA from wildlife samples was carried out. Partial mitochondrion DNA 12S rRNA and Cyt b genes were amplified and sequenced. The original animal identifications of different wildlife samples were determined by BLAST search of the sequences in GenBank. To confirm the results of BLAST, genetic distance and phylogenetic tree were analyzed by MEGA 7.0. Result The genome DNA was successfully extracted from wildlife samples and partial mitochondrion DNA 12S rRNA and Cyt b genes were amplified. In the identification of Ursus thibetanus, Pantholops hodgsonii, Capricornis milneedwardsii, and Manis javanica sample species, both 12S rRNA gene marker and Cyt b gene marker can identify the species of wild animals successfully. However, in the identification of Capra ibex, unlike Cyt b gene marker, 12S rRNA gene marker cannot distinguish Capra ibex from Capra hircus gene sequences. Phylogenetic trees constructed based on 12S rRNA and Cyt b sequences were consistent with BLAST analysis. Conclusion The identification ability of single locus marker for some species is poor, thus it is recommended to identify wild animal species by selecting more than two gene marker loci.

Key words: wild animal, species identification, 12S rRNA, Cyt b, gene marker

中图分类号: 

  • Q959