Chines Journal of Vector Biology and Control ›› 2015, Vol. 26 ›› Issue (3): 286-289.DOI: 10.11853/j.issn.1003.4692.2015.03.017

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Comparison of four DNA barcoding in Rattus losea identification

HU Qun1, MA Si-jie1, QIU Jiong-liang2   

  1. 1 Daxie Entry-Exit Inspection and Quarantine Bureau, Ningbo 315812, Zhejiang Province, China;
    2 Ningbo Entry-Exit Inspection and Quarantine Bureau
  • Received:2014-12-10 Online:2015-06-20 Published:2015-06-20
  • Supported by:

    Supported by the Ningbo Natural Science Fund(No. 2013A610240)

4种DNA条形码在黄毛鼠种类鉴定中的比较

胡群1, 马思杰1, 裘炯良2   

  1. 1 大榭出入境检验检疫局鼠传疾病检测实验室, 浙江宁波315812;
    2 宁波出入境检验检疫局
  • 作者简介:胡群,男,硕士,高级工程师,主要从事口岸医学媒介鉴定与病原体检测研究,Email: nbhuqun@sina.com
  • 基金资助:

    宁波市自然科学基金(2013A610240)

Abstract:

Objective To evaluate CO Ⅰ, Cytb, 16S rRNA, D-loop genes as DNA barcodes for Rattus losea species identification. Methods Genome DNA was extracted from a R. losea sample HX41 captured in Daxie port. The mitochondrial COⅠ, Cytb, 16S rRNA, D-loop genes were amplified using the specific primers and sequenced. The phylogenetic trees was constructed based on the nucleotide sequences of HX41 with Rattus published in GenBank and the homology was analyzed. Results HX41 was located in the branch of the R. losea on phylogenetic tree based on COⅠ, Cytb, and D-loop genes, which form a single branch. The genetic distance analysis showed that the COⅠ gene distance of HX41 to R. losea HN158 is nearest, the value is 0.0034. The Cytb gene distance of HX41 to R. tanezumi HN99 is nearest, the value is 0.0355, but the species database of HN99 in GenBank was incorrect. The D-loop gene of HX41 to R. losea HN105 is nearest, the value is 0.0439. Because of lacking the data of R. losea 16S rRNA gene in the GenBank, the phylogenetic tree cannot be constructed. Conclusion Using molecular identification method for rat species identification, it is necessary to choose suitable DNA barcode according to the integrity and accuracy of the GenBank database.

Key words: Rattus losea, Species identification, DNA barcoding

摘要:

目的 比较COⅠ、Cytb、16S rRNA、D-loop四个DNA条形码对黄毛鼠鼠种的鉴定效果。方法 将大榭港区捕获的1只黄毛鼠样本HX41,提取基因组DNA,使用特异性引物对线粒体COⅠ、Cytb、16S rRNA、D-loop基因进行扩增并对扩增产物进行测序。将测序结果与GenBank中的相似鼠种DNA构建分子进化树进行同源性分析。结果 以COⅠ、Cytb、D-loop基因所构建分子进化树中,黄毛鼠均能独立形成分支,且HX41均处于黄毛鼠分支上。HX41的COⅠ基因序列与黄毛鼠HN158 株遗传距离最为接近,值为0.0034;Cytb基因序列与黄胸鼠HN99株遗传距离最为接近,值为0.0355,但HN99鼠种数据存在错误;D-loop基因序列与黄毛鼠HN105株遗传距离最为接近,值为0.0439。由于现有GenBank中缺少黄毛鼠16S rRNA基因序列,无法构建进化树。结论 对鼠种进行分子鉴定时,有必要根据GenBank数据库的完整性和准确性选择合适的DNA条形码。

关键词: 黄毛鼠, 种类鉴定, DNA条形码

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