藏药羌活鱼的COⅠ分子鉴定  

Molecular identification of Tibetan medicine Qianghuoyu by COⅠ

作　　者：杜文杰[1] 刘海青 徐婧[1] 骆桂法 梅之南[1] 

机构地区：[1]中南民族大学药学院,湖北武汉430074 [2]青海省食品药品检验所,青海西宁810016

出　　处：《中国中药杂志》2015年第3期395-398,共4页China Journal of Chinese Materia Medica

基　　金：国家药典委员会民族药材质量标准研究项目(M13)

摘　　要：对动物线粒体基因COⅠ进行PCR扩增及双向测序,得到羌活鱼及其混伪品无斑肥源和截趾虎的COⅠ基因序列。通过序列分析比对,根据序列差异设计特异性引物SJYW1,SJYW2,改变退火温度及循环数以得到特异性的反应条件。结果表明,当复性温度为54℃,25个循环时,正品羌活鱼在约350 bp处有明显扩增带,而无斑肥源和截趾虎没有扩增带。该实验具有较高的特异性、重复性,可用于羌活鱼的鉴别。The CO I gene sequences of Qianghuoyu, Pachytriton labiatus and Gehyra mutilata were achieved by PCR amplification and hi-directional sequencing. Furthermore, a pair of specific primers SJYWI and SJYW2 in the non-conservative district were de- signed through sequence alignment. The PCR reaction condition was established by changing the annealing temperature and cycle num- bers. The results showed that 350 bp DNA fragment was amplified from Qianghuoyu in PCR with annealed temperature at 54℃ and the cycle number was 25 cycles, whereas not any DNA fragment was amplified from P. labiatus and G. mutilata under the same reaction condition. This method is well-performed in the identification of Qianghuoyu for its excellent specificity and repeatability.

关 键 词：羌活鱼 山溪鲵 混伪品 PCR 特异性 分子鉴定 

分 类 号：R29[医药卫生—民族医学]