DNA条形码技术在动物类药材熊胆粉及其混伪品鉴定中的应用  

Identification of bear bile powder and its adulterants by using DNA barcoding technology

作　　者：许亚春 熊超[2] 姜春丽[1,2] 段永波[1] 孙伟[2] 刘绍勇 薛东升 薛建平[1] XU Ya-chun;XIONG Chao;JIANG Chun-lil,;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;Shanghai Kaibao Pharmaceutical Co. , Ltd. , Shanghai 201401, China)

机构地区：[1]淮北师范大学生命科学学院,淮北235000 [2]中国中医科学院中药研究所,北京100700 [3]上海凯宝药业股份有限公司,上海201401

出　　处：《中国中药杂志》2018年第4期645-650,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81573518,81603248);安徽省自然科学基金项目(1408085MC58);中央级公益性科研院所基本科研业务费专项(GH2017-01-01,GH2017-01)

摘　　要：应用DNA条形码技术对名贵中药材熊胆粉及混伪品进行DNA条形码鉴定研究，并建立其标准实验流程，以保障熊胆粉的安全有效利用。对收集到的12份熊胆粉样品进行DNA提取、PCR（聚合酶链式反应）扩增并双向测序、运用CodonCodeAlignerV7．0．1进行序列剪切去除引物区得到CO／序列，同时与GenBank中得到棕熊及混伪品COI序列，运用MEGA7．0对研究的5个物种50条序列比对分析，计算变异位点及种内和种问K2P（Kimura2-Parameter）遗传距离并构建邻接（NJ）树。黑熊、棕熊的种内最大K2P遗传距离远小于其混伪品间最小K2P遗传距离；NJ树结果显示黑熊、棕熊各聚为一支，均可以与混伪品明显区分开。DNA条形码技术为一种安全便捷可靠的物种鉴定技术，建立动物类药材COI序列标准话流程，在中药研究领域具有重要的作用。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 thi- betanus for DNA extraction, PCR （polymerase chain reaction） and sequence, then using CodonCode Aligner V 7. 0. 1 shear primer re- gion to obtain CO1 sequence. The CO1 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 con- strutting the Neighbor-joining tree （NJ）. The results showed that the maximum K2P genetic distance of bear bile powder of U. thibeta- nus 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 CO1 sequences for animal medicines.

关 键 词：熊胆粉 COI序列 DNA条形码技术 分子鉴定 

分 类 号：R282.5[医药卫生—中药学]