中国中药杂志

2019, v.44(15) 3261-3267

[打印本页] [关闭]
本期目录(Current Issue) | 过刊浏览(Past Issue) | 高级检索(Advanced Search)

位点特异性PCR鉴别延胡索、齿瓣延胡索和夏天无药材
Identification of Corydalis yanhusuo,C. turtschaninovii,C. decumbens by allele-specific PCR

产清云;蒋露;程铭恩;储姗姗;于大庆;谢晋;查良平;彭华胜;
CHAN Qing-yun;JIANG Lu;CHENG Ming-en;CHU Shan-shan;YU Da-qing;XIE Jin;ZHA Liang-ping;PENG Hua-sheng;School of Pharmacy,Anhui University of Chinese Medicine;School of Pharmaceutical Sciences,Anhui Medical University;Institute of Conservation and Development of Traditional Chinese Medicine Resources,Anhui Academy of Chinese Medicine;

摘要(Abstract):

建立一种鉴别延胡索、齿瓣延胡索和夏天无的DNA分子标记鉴别方法。通过测序获得延胡索、齿瓣延胡索、夏天无及其近缘种14种共56个样品的mat K,trn G和trn H-psb A序列,利用Bio Edit 7. 2. 2软件比对获得SNP位点,根据变异位点设计AS-PCR鉴别引物,并优化PCR条件,根据特异性条带分别对延胡索、齿瓣延胡索和夏天无进行鉴定。结果显示模板量(0. 6~1 200 ng)和退火温度(42~60℃)对扩增结果影响较小,循环次数对扩增结果影响较大。在循环数为20时,延胡索、齿瓣延胡索和夏天无分别扩出297 bp(mat K)、353 bp(trn G)和544 bp(mat K)的特异性条带。该研究所建立的方法对延胡索的最低检测限为6 ng,对夏天无的最低检测限为60 ng,齿瓣延胡索低于0. 6 ng。该文所建立的位点特异性PCR法能特异性鉴别延胡索、齿瓣延胡索和夏天无。
To establish a DNA molecular markers method for identification of Corydalis yanhusuo,C. turtschaninovii and C. decumbens,the mat K,trn G and psb A-trn H sequences of 56 samples from 14 species of C. yanhusuo,C. turtschaninovii,C. decumbens and their related species were obtained by sequencing. The SNP loci were obtained by Bio Edit 7. 2. 2 software. The primers for AS-PCR identification were designed based on the mutation sites,and the conditions of PCR were optimized to identify C. yanhusuo,C. turtschaninovii,and C. decumbens according to the specific bands. The results showed that the amount of template( 0. 6-1 200 ng)and annealing temperature( 42-60 ℃) had little influence on the amplification results,and the number of cycles had much influence on the amplification results. When the number of cycles was 20,the specific bands of 297 bp( mat K),353 bp( trn G) and 544 bp( mat K) were amplified from C. yanhusuo,C. turtschaninovii and C. decumbens,respectively. The method established in this study had a minimum detection limit of 6 ng for C. yanhusuo,60 ng for C. decumbens and less than 0. 6 ng for C. turtschaninovii. Thus,the allelespecific PCR method established in the research can specifically identify C. yanhusuo,C. turtschaninovii,and C. decumbens.

关键词(KeyWords): 延胡索;齿瓣延胡索;夏天无;特异性PCR;分子鉴定
Corydalis yanhusuo;Corydalis turtschaninovii;Corydalis decumbens;specific polymerase chain reaction;molecular identification

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81703633,81803675)

作者(Author): 产清云;蒋露;程铭恩;储姗姗;于大庆;谢晋;查良平;彭华胜;
CHAN Qing-yun;JIANG Lu;CHENG Ming-en;CHU Shan-shan;YU Da-qing;XIE Jin;ZHA Liang-ping;PENG Hua-sheng;School of Pharmacy,Anhui University of Chinese Medicine;School of Pharmaceutical Sciences,Anhui Medical University;Institute of Conservation and Development of Traditional Chinese Medicine Resources,Anhui Academy of Chinese Medicine;

Email:

DOI:

参考文献(References):

扩展功能
本文信息
服务与反馈
本文关键词相关文章
本文作者相关文章
中国知网
分享