Xu Ya-Chun, Xiong Chao, Jiang Chun-Li, Duan Yong-Bo, Sun Wei, Liu Shao-Yong, Xue Dong-Sheng, Xue Jian-Ping
School of Life Sciences, Huaibei Normal University, Huaibei 235000, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2018 Feb;43(4):645-650. doi: 10.19540/j.cnki.cjcmm.2018.0014.
To identify the precious bile powder and its adulterants by DNA barcoding, and establish its standard experimental process to ensure the safe and effective utilization. Total twelve sequences from samples of bear bile powder which come from Ursus thibetanus for DNA extraction, PCR(polymerase chain reaction) and sequence, then using CodonCode Aligner V 7.0.1 shear primer region to obtain COI sequence. The COI sequences of U. arctos and their adulterants were obtained from GenBank. MEGA7.0 software was applied for analyzing mutation, calculating intraspecific and interspecific K2P(Kimura 2-Parameter) genetic distance and constructing the Neighbor-joining tree(NJ). The results showed that the maximum K2P genetic distance of bear bile powder of U. thibetanus and U. arctos are far less than minimum K2P genetic distance within its adulterants species, and the results of NJ tree demonstrated that each species could be distinguished from the counterfeits obviously. DNA barcoding is a safe, convenient and reliable technique for species identification, and it is important to establish the standard sequence of COI sequences for animal medicines.
通过DNA条形码技术鉴别珍贵的熊胆粉及其掺伪品，建立其标准实验流程以确保安全有效利用。对采自黑熊的12份熊胆粉样本进行DNA提取、PCR（聚合酶链反应）及测序，然后使用CodonCode Aligner V 7.0.1剪切引物区域以获得COI序列。从GenBank获取棕熊及其掺伪品的COI序列。应用MEGA7.0软件分析突变，计算种内和种间的K2P（Kimura双参数）遗传距离并构建邻接树（NJ）。结果表明，黑熊和棕熊熊胆粉的最大K2P遗传距离远小于其掺伪品物种内的最小K2P遗传距离，NJ树结果表明各物种可明显与假冒品区分开来。DNA条形码技术是一种安全、便捷、可靠的物种鉴定技术，建立动物药COI序列的标准序列很重要。
