[关键词]
[摘要]
目的 本研究旨在建立一种简便、实用的鳖甲DNA分子鉴定方法。 方法 根据中华鳖和鳖甲混淆品原动物的12SrRNA基因片段的序列数据库, 找出中华鳖与其它鳖科动物有明显区别的位点, 设计1对能特异性鉴别中华鳖的引物, 然后利用鳖甲中残存的DNA, 通过PCR扩增, 即可准确鉴别鳖甲。 结果 用这对引物对不同来源的10块鳖甲进行鉴定结果表明, 其中有3块为伪品, 结果与DNA序列分析鉴定一致。 还测定了斑鼋和鳖甲一伪品12SrRNA基因片段序列。结论 该引物配置药材鉴定试剂盒可在鳖甲类药材鉴定使用。
[Key word]
[Abstract]
Object To develop a convenient and practical method for the identification of Carapax Trionycis.Methods Based on the sequence variations of 12SrRNA gene between Pelodiscus sinensis and other softshell turtles, a pair of allele-specific primers was designed to distinguish P. sinensis from other species of Trionychidae. DNA were extracted and anplified and Carapax Trionycis could be identified accu-rately by polymerase chain reaction(PCR) using the primers. Results Ten samples of turtle shell from different sources were indentified by the allele-specific PCR with the primers. The result indicated that three samples were substitutes of Carapax Trionycis, consilient with the result from DNA sequence analy-sis. The mitochondrial 12SrRNA gene fragment of P. maculatus and a faked imitation had also been se-quenced. Conclusion The primers could be used as key components in Carapax Trionycis identification kit.
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[基金项目]
国家自然科学基金(No.39870913)、江苏教委科学基金(98KJB360010)和江苏省“333工程”人才培养基金资助