中国中药杂志

2013, v.38(16) 2563-2566

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断肠草与金银花类药材水提液特异性PCR鉴定方法研究
Study on molecular identification of water extracts from Gelsemium elegans and Lonicera japonica and its close species by specific PCR amplification

崔占虎;蒋超;黄璐琦;李旻辉;周涛;周立社;袁媛;
CUI Zhan-hu1,2,JIANG Chao1,HUANG Lu-qi1,LI Min-hui2,ZHOU Tao3,ZHOU Li-she2,YUAN Yuan1*(1.National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China; 2.Inner Mongolia University of Science and Technology,Baotou Medical College,Baotou 014060,China; 3.Guiyang Collage of Traditional Chinese Medicine,Guiyang 550002,China)

摘要(Abstract):

目的:研究断肠草与金银花类药材水提液的DNA分子鉴别方法。方法:采集不同产地的断肠草药材13份、金银花32份、山银花21份和水银花5份,所有样品提取总DNA,并对断肠草、金银花、山银花的水提液进行DNA提取。通过对断肠草ps-bA-trnH片段进行扩增、测序,并搜索GenBank数据库中收录的中金银花类药材基原植物psbA-trnH序列,进行同源比对后根据其变异位点设计特异性鉴别引物。结果:通过使用特异性引物对所有样品进行PCR扩增,发现断肠草可扩增出97 bp片段,而金银花类药材则不能扩增出条带。结论:通过特异性PCR的方法实现了断肠草与金银花类药材水提液的快速、准确鉴别。
Objective: To explore the new method of discriminating Gelsemium elegans from Lonicera japonica and its close species by using specific PCR amplification.Method: Thirteen samples of the different G.elegans materials and 58 samples of L.japonica,L.macranthoides and L.dasystyla were collected.The total DNA of the samples were extracted,and the DNA of G.elegans,L.japonica and L.macranthoides water extracts were extracted.PsbA-trnH sequence from G.elegans was amplified by PCR and sequenced unidirectionally,ClustulW was used to align psbA-trnH sequences of the G.elegans and L.japonica and its close species from GenBank database.Result: All samples were amplified by PCR with specific primer,DNA from G.elegans would be amplified 97 bp whereas PCR products from all of Lonicera samples had not bands.Conclusion: Specific PCR amplification can be used to identify G.elegans from L.japonica and its close species successfully and is an efficient molecular marker for authentication of G.elegans and L.japonica and its close species.

关键词(KeyWords): 断肠草;水提液;特异性PCR;分子鉴定
Gelsemium elegans;water extracts;specific PCR amplification;molecular identification

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基金项目(Foundation): 北京市科技专项“道地中药功能基因组北京市重点实验室2012年阶梯计划项目”;; 贵州省中药现代化科技产业研究开发专项(黔科合ZY字[2013]3001号);; 公安部重点研究计划项目(2013012DYJD07)

作者(Author): 崔占虎;蒋超;黄璐琦;李旻辉;周涛;周立社;袁媛;
CUI Zhan-hu1,2,JIANG Chao1,HUANG Lu-qi1,LI Min-hui2,ZHOU Tao3,ZHOU Li-she2,YUAN Yuan1*(1.National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China; 2.Inner Mongolia University of Science and Technology,Baotou Medical College,Baotou 014060,China; 3.Guiyang Collage of Traditional Chinese Medicine,Guiyang 550002,China)

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