基于COI序列哈蟆油基原动物DNA条形码鉴定研究.docVIP

基于COI序列哈蟆油基原动物DNA条形码鉴定研究 　　[摘要] 哈蟆油具有较高的经济和社会价值，其混伪品较多，给市场造成了很大的混乱。利用基于COI序列的DNA条形码技术对已鉴定的东北林蛙、中国林蛙、桓仁林蛙和黑龙江林蛙进行PCR扩增和测序，建立4种林蛙COI基因数据库；将所测的序列与GenBank上的序列进行比对，显示为上述4种林蛙；应用MEGA（molecular evolutionary genetics analysis）7.0软件计算K2P遗传距离并构建NJ（neighbor-joining）系统聚类树，所测4种林蛙的序列和GenBank上下载的4种林蛙的序列分别聚为1支，但与GenBank上下载的2个外群独立分开，东北林蛙的COI基因可能存在地域性差异，但该技术并不能鉴别林蛙的雌雄。结果显示DNA条形码技术能够准确鉴定哈蟆油的基原动物，说明DNA条形码COI为哈蟆油基原动物的鉴定提供一个新的手段和方法。 　　[关键词] 哈蟆油；东北林蛙；中国林蛙；DNA条形码；鉴定 　　Identification of Ranae Oviductus original animal based on COI sequences 　　WANG Meng-hu，KANG Ting-guo*，XU Liang*，YANG Yan-yun，CAI Zhen-jiao 　　（Liaoning University of Traditional Chinese Medicine，Dalian 116600，China） 　　[Abstract] 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. 　　[Keywords] Ranae Oviductus； Rana dybowskii； Rana chensinensis； DNA barcode； identify