Wang Meng-Hu, Kang Ting-Guo, Xu Liang, Yang Yan-Yun, Cai Zhen-Jiao
Liaoning University of Traditional Chinese Medicine, Dalian 116600, China.
Zhongguo Zhong Yao Za Zhi. 2017 Apr;42(8):1572-1577. doi: 10.19540/j.cnki.cjcmm.20170223.016.
Ranae Oviductus has a high economic and social value, but its adulterants are more numerous, which causes a great confusion to the market. Using DNA bar code technology based on COI sequence for PCR amplification and sequencing of the identified Rana dybowskii, R. chensinensis, R. huanrensis and R. amurensiss, the COI gene database of four species of Rana was established, and comparing the measured sequence with the sequence of GenBank, four kinds of Rana were identified. The MEGA (molecular evolutionary genetics analysis) 7 .0 software was used to calculate the genetic distance of K2P and construct the NJ (neighbor-joining) system cluster tree. The sequence of the four species of Rana measured were clustered into one group with the sequence of the four kinds of Rana downloaded from GenBank, but separated from the two outer groups downloaded from GenBank. The COI gene of the R. dybowskii was likely to have regional differences, however this technique failed to distinguish male and female Rana. The results showed that DNA bar code technology could accurately identify the base of original animal of R. oviductus. It indicates that DNA bar code COI provides a new method for the identification of R. oviductus.
林蛙油具有很高的经济和社会价值,但其掺伪品众多,给市场造成极大混乱。利用基于COI序列的DNA条形码技术对鉴定出的东北林蛙、中国林蛙、桓仁林蛙和黑龙江林蛙进行PCR扩增和测序,建立了4种林蛙的COI基因数据库,并将测得序列与GenBank序列进行比对,鉴定出4种林蛙。使用MEGA(分子进化遗传学分析)7.0软件计算K2P遗传距离并构建NJ(邻接法)系统聚类树。测得的4种林蛙序列与从GenBank下载的4种林蛙序列聚类为一组,但与从GenBank下载的另外两组分离。东北林蛙的COI基因可能存在区域差异,然而该技术未能区分林蛙的雌雄。结果表明,DNA条形码技术能够准确鉴定林蛙油的原动物基源。这表明DNA条形码COI为林蛙油的鉴定提供了一种新方法。