中国中药杂志

2013, v.38(16) 2581-2585

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基于位点特异性PCR的黄芪与红芪鉴别方法研究
Study on identification of Astragali Radix and Hedysari Radix by PCR amplification of specific alleles

龙平;崔占虎;李虔全;徐建平;张春红;周立社;李旻辉;
LONG Ping,CUI Zhan-hu,LI Qian-quan,Xu Jian-ping,ZHANG Chun-hong,ZHOU Li-she*,LI Min-hui*(Baotou Medical College,Inner Mongolia,Baotou 014060,China)

摘要(Abstract):

研究黄芪与红芪药材之间的DNA分子鉴别方法。采集不同产地的黄芪药材30份、红芪28份,所有样品进行总DNA的提取,通过对黄芪及红芪药材trnL-trnF片段进行扩增、测序,进行同源比对后根据其变异位点设计特异性鉴别引物。并对位点特异性PCR方法进行条件优化。通过使用特异性引物对所有样品进行PCR扩增,发现黄芪可扩增出136 bp片段,红芪可以扩增出323 bp片段。黄芪中掺入10%以上红芪可以扩增出红芪特异鉴别条带。通过位点特异性PCR的方法实现了黄芪与红芪药材的快速、准确鉴别。
To explore the new method of discriminating Astragali Radix and Hedysari Radix by using PCR amplification of specific alleles,30 samples of the different Astragali Radix materials and 28 samples of Hedysari Radix were collected.The total DNA of all samples were extracted,trnL-trnF sequence from Astragali Radix and Hedysari Radix was amplified by PCR and sequenced unidirectionally.These sequences were aligned by using Clustul W.Primer was designed and the PCR reaction systems including annealing temperature,dNTP,etc were optimized.All samples were amplified by PCR with specific primer,DNA from Astragali Radix would be amplified 136 bp,whereas PCR products from all of Hedysari Radix were 323 bp.This method can detect 10% of intentional Hedysari Radix DNA into Astragali Radix.PCR amplification of alleles can be used to identify Astragali Radix and Hedysari Radix successfully and is an efficient molecular marker for authentication of Astragali Radix and Hedysari Radix.

关键词(KeyWords): 黄芪;位点特异性PCR;分子鉴定
Astragali Radix;PCR amplification of specific alleles;molecular identification

Abstract:

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基金项目(Foundation): 国家“十二五”科技支撑计划项目(2012BA128B02);; 中医药行业科研专项(201107009);; 包头科技发展项目(2011X1002)

作者(Author): 龙平;崔占虎;李虔全;徐建平;张春红;周立社;李旻辉;
LONG Ping,CUI Zhan-hu,LI Qian-quan,Xu Jian-ping,ZHANG Chun-hong,ZHOU Li-she*,LI Min-hui*(Baotou Medical College,Inner Mongolia,Baotou 014060,China)

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