Wu P, Zhou K, Xu L, Teng J
Department of Biology, Nanjing Normal University, Nanjing 210097.
Yao Xue Xue Bao. 1998;33(4):304-9.
This paper reports a new method to identify the Chinese drug turtle shells using PCR product direct sequencing method. DNA was extracted from tissues of the Chinese three-keeled pond turtle Chinemys reevesii and 20 other species of turtles occurring in China and Southeast Asian countries. One hundred and ten base pairs of mitochondrial 12S rRNA gene fragment were amplified from the extract using PCR technique and obtained sequences. These sequences were used to construct 12S rRNA gene fragment sequence database for the 21 turtle species. Comparison of these sequences indicated that the sequence from the Chinese three-keeled pond turtle is different from that of all the other 20 turtle species. The sequence divergence is 3.7-15.7%. DNA was extracted from 0.1-0.5 g of shell from 19 turtle shells provided by the Jiangsu Institute for Drug Control and 12S rRNA gene fragment was amplified and sequenced. Comparison of the sequences from the 19 turtle shells and 12S rRNA gene fragment sequence database indicated that only 3 samples are shells of the Chinese three-keeled pond turtle specified in the Phamacopoeia of the People's Republic of China and the others are substitutes. The technique used in the present paper was found to be effective and reliable for the identification of turtle shells.
本文报道了一种采用PCR产物直接测序法鉴定中药材龟甲的新方法。从中国中华鳖及中国和东南亚国家的其他20种龟类的组织中提取DNA。利用PCR技术从提取物中扩增出110个碱基对的线粒体12S rRNA基因片段并获得序列。这些序列用于构建21种龟类的12S rRNA基因片段序列数据库。这些序列的比较表明，中华鳖的序列与其他20种龟类的序列不同。序列差异为3.7%-15.7%。从江苏省药品检验所提供的19个龟甲中提取0.1-0.5g龟甲的DNA，扩增并测序12S rRNA基因片段。将19个龟甲的序列与12S rRNA基因片段序列数据库进行比较，结果表明，只有3个样品是《中华人民共和国药典》规定的中华鳖龟甲，其他均为替代品。本文所采用的技术被发现对龟甲的鉴定是有效和可靠的。
