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本研究以常用于动物种属鉴定的Cytb基因位点为研究对象,利用所测得的30种盗猎案件中常见的野生动物Cytb基因部分片段序列及NCBI数据库中下载的该物种序列及其近缘物种序列,构建系统进化树。根据进化树的聚类情况,判断NCBI数据库中的相关基因序列或物种名称的正确性,并对其中错误序列的登陆号进行标记,以防对后续涉案动物的准确鉴定造成影响。分别从30种常见涉案野生动物(共35份样本)中提取线粒体DNA,并利用通用引物扩增线粒体DNA上的Cytb基因部分片段并进行测序分析。通过NCBI数据库的Blast比对功能,筛选出与本研究物种同源性由高到低的物种,并从NCBI基因数据库中下载此类近缘物种的Cytb基因序列共482条,利用MEGA软件构建该物种的系统进化树。比对发现NCBI中登录号为DQ246798、KP202269、AY286434等3个序列所对应物种拉丁名错误。登录号为AY509634、AJ131639、AM072747等5个序列所对应物种拉丁名可能存在疑问。NCBI数据库数据可靠性有待进一步验证,只能作为涉案物种鉴定的参考数据之一,可借助构建系统进化树等方法来确认其结果的准确性。
Abstract：In this research, the Cytb gene site, which is often used in animal species identificantion, was used as the research object. Through combination of directly sequencing of 30 kinds of wild animal species that were routinely encountered in illegal trade as well as their close species and downloading reference sequence from NCBI database for the Cytb gene fragment, phylogenetic tree was constructed. According to the resulting evolutionary relationship,reliability of gene sequences or species naming in NCBI database was evaluated. Moreover, accession numbers of related error sequence were marked to prevent the impact on the accurate identification of the animals involved in the subsequent case. Mitochondrial DNA was extracted from 30 kinds of wild animal species(a total of 35 samples), and the Cytb gene fragment of mitochondrial DNA was amplified by universal primers and then sequenced. Through comparison function built in Blast module, the sequences of the same or close species of the tested species(total482) were downloaded from the NCBI database. MEGA software was used to construct phylogenetic tree of the species. By comparison, we found that the Latin names of three species with corresponding register numbers including DQ246798, KP202269, and AY286434 were wrong in NCBI. There were some doubts about the Latin name of five species with corresponding login numbers, including AY509634, AJ131639, AM072747 and so on.The reliability of NCBI database needs to be further verified, and can only be used as one of the reference for species identification. The phylogenetic tree can be added to confirm the accuracy of the result.
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基本信息:
DOI：10.13417/j.gab.038.003981
中图分类号:Q78
引用信息:
[1]黄娅琳,徐燕红,侯森林,等.基于Cytb基因的NCBI数据库序列可靠性分析[J].基因组学与应用生物学,2019,38(09):3981-3986.DOI:10.13417/j.gab.038.003981.
基金信息:
中央高校基本科研业务费专项资金项目(LGZD201707)和预研项目(LGY201601)共同资助
2019-09-25
2019-09-25