中国中药杂志

2013, v.38(16) 2567-2570

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白术、苍术种子位点特异性PCR鉴别方法研究
Authenticate of Atractylodes macrocephala seed by amplification refractory mutation system

曹亮;蒋超;彭华胜;陈敏;袁媛;
CAO Liang1,2,JIANG Chao1,PENG Hua-sheng3,CHEN Min1,YUAN Yuan1*(1.National Resource Center for Chinese Materia Medica,China Academy of Chinese Medicinal Sciences,Beijing 100700,China; 2.Institute of Agricultural and Biology Resource Utilization,Hunan Academy of Agricultural Sciences,Changsha 410125,China; 3.Anhui College of Traditional Chinese Medicine Chinese,Hefei 230038,China)

摘要(Abstract):

目的:筛选白术、苍术鉴别SNP位点,并设计特异性引物,实现白术、苍术种子的快速鉴定。方法:通过测序及GenBank中下载获得白术、苍术的psbA-trnH序列,通过使用Bioedit软件比对获得SNP位点,设计位点特异性PCR引物,将ITS通用引物加入到特异性引物构建多重PCR体系,并优化PCR条件,对白术、苍术种子进行分子鉴定。结果:构建了多重PCR鉴别体系,通过一个PCR反应,能扩增出苍术、白术约643 bp的ITS DNA质控条带,扩增出白术172 bp的特异性鉴别条带,实现白术、苍术鉴别。结论:使用位点特异性PCR技术可以有效鉴别白术、苍术种子。
Objective: To design specific primers and authenticate Atractylodes macrocephala from Atractylodes lancea and A.chinensis.Method: SNPs in the psbA-trnH sequences of Atractylodes were found by ClustulW program and Bioedit software.Primers for authentic A.macrocephala is designed according to the SNP site,and ITS sequence universal primers plus to the authentic primer to construct a multi-PCR reaction system,and then optimized the PCR reaction system.Result: 172 bp band special for A.macrocephala were found using multi-PCR reaction.Conclusion: The multi-PCR reaction system could be applied to identify A.macrocephala seed.

关键词(KeyWords): 白术;SNP;分子鉴定;种子
Atractylodes macrocephala;SNP;molecular identification;seed

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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”

作者(Author): 曹亮;蒋超;彭华胜;陈敏;袁媛;
CAO Liang1,2,JIANG Chao1,PENG Hua-sheng3,CHEN Min1,YUAN Yuan1*(1.National Resource Center for Chinese Materia Medica,China Academy of Chinese Medicinal Sciences,Beijing 100700,China; 2.Institute of Agricultural and Biology Resource Utilization,Hunan Academy of Agricultural Sciences,Changsha 410125,China; 3.Anhui College of Traditional Chinese Medicine Chinese,Hefei 230038,China)

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