DNA条形码技术在动物角制品鉴别中的应用  

Application of DNA Barcoding in Identification of Animal Horn Products

作　　者：王殿夫 刘洪伟 巴哈提古丽·马那提拜 麻丽丹 WANG Dian-fu;LIU Hong-wei;Manatibai Guhatiguli;MA Li-dan(Office of Academic Research,Eastern Liaoning University,Dandong 118001,China;Dandong Customs,Dandong 118000,China;Urumqi Customs,Urumqi 830000,China)

机构地区：[1]辽东学院科研处,辽宁丹东118001 [2]丹东海关,辽宁丹东118000 [3]新疆海关,新疆乌鲁木齐830000

出　　处：《辽东学院学报（自然科学版）》2021年第2期84-90,共7页Journal of Eastern Liaoning University:Natural Science Edition

基　　金：海关总署科研项目(2019HK040)。

摘　　要：利用动物线粒体12S rRNA和16S rRNA基因序列片段对动物角制品进行种属鉴别,为DNA条形码技术在角制品分子鉴定提供有效方法。收集犀牛角、水牛角、黄牛角和山羊角作为研究材料,采用优化提取步骤的商业化提取试剂盒进行基因组DNA的提取,以提高基因组DNA的浓度及质量。采用PCR扩增及测序技术获得所有样本12S rRNA和16S rRNA基因部分序列,使用Blast程序分别进行序列比对分析,通过下载Genbank相关物种的12S rRNA和16S rRNA序列,分别比较两个基因片段种内和种间距离,分别构建系统进化树。结果显示,12S rRNA序列所有样本扩增成功,16S rRNA序列仅犀牛角和黄牛角扩增成功。遗传距离分析显示各检材种间差异均大于种内差异,收集检材在12S rRNA和16S rRNA引物构建的进化树中表现出良好的单系性和较高的置信度。表明12S rRNA基因片段的DNA条形码技术对动物角制品及其它混伪品具有更好的鉴别能力,可提高物种鉴定的准确性和灵敏度。Animal mitochondria 12S rRNA and 16S rRNA gene sequences fragment was used to identify the animal horn products,by which to explore an effective method for molecular identification of horn products by DNA barcoding.The genomic DNA of rhino horn,buffalo horn,ox horn and goat horn were extracted with a commercial extraction kit.The 12S rRNA and 16S rRNA sequences of GenBank related species were downloaded and the intra-specific and inter-specific genetic distances of them were compared and phylogenetic trees were constructed for the two gene fragments.It was found that the 12S rRNA sequence was amplified successfully with all samples while the 16S rRNA sequence was amplified only in rhino horn and cattle horn.Genetic distance analysis indicated that,the difference between species was greater than that within species and the collected samples showed good monophyletic and high confidence in the phylogenetic tree constructed by 12S rRNA and 16S rRNA primers.It is concluded that DNA barcoding of 12S rRNA gene fragments is more reliable in identifying animal horn products and other counterfeits,which can improve the accuracy and sensitivity of species identification.

关 键 词：动物角制品 DNA条形码技术 动物线粒体 

分 类 号：Q523[生物学—生物化学]