目的 以河北省鼠疫疫源地常见鼠为对象,探讨DNA条形码技术在鼠类鉴定方面的可行性。方法 2012年在河北省鼠疫疫源地内采集鼠肝脏标本,并保存整只鼠,制作剥制标本,提取基因组DNA,用通用引物PCR 法扩增线粒体COⅠ基因,并测序。将测序结果与GenBank中其他鼠类物种的DNA条形码进行BLAST 比对,并构建分子进化树。结果87份样本均能通过PCR 扩增出特异性COⅠ基因条带,其中85份样本所构建的分子进化树结果与形态学鉴定结果一致,2份样本结果有差异,经反复鉴定头骨及形态标本,发现为现场鉴定错误。结论 DNA条形码技术能够对鼠类进行有效的物种鉴定,同时还可以探讨啮齿动物属、种分类单元的系统发育问题。
闫东,鲁亮,金圣浩,史献明,崔耀仁,刘冠纯,李玉伟,刘起勇,郑楠,康东梅,白雪薇,牛艳芬,陈永明,周松,兰晓宇,李振海
. DNA条形码技术在河北省鼠疫疫源地鼠种鉴定中的应用 [J]. 中国媒介生物学及控制杂志, 2014
, 25(1)
: 21
-23
.
DOI: 10.11853/j.issn.1003.4692.2014.01.006
Objective To evaluate the feasibility of DNA barcoding in identifying the rodent species based on the common rodents in the natural focus of plague in Hebei province, China. Methods The rodent liver specimens were collected in the natural focus of plague in Hebei in 2012; the whole rodents were conserved, and then the stuffed specimens were made; genomic DNA was extracted from the rodent liver. The mitochondrial COⅠ gene was amplified by consensus primer PCR and then sequenced. BLAST comparison was made between the sequencing results and the DNA bar codes of other rodent species in GenBank, and molecular evolutionary tree was constructed. Results In this research, specific COⅠ gene bands were obtained by PCR amplification in all 87 samples. Among all samples, 85 had the molecular evolutionary tree in accordance with the results of morphological identification, and 2 had different results, but the field identification was found to be incorrect by identifying the skulls and morphological specimens repeatedly. Conclusion DNA barcoding technique can be used to effectively identify the rodent species. At the same time, it can also be used for discussing the phylogenetic relationship of rodent genus and species classification.
