威灵仙的聚合酶链反应-单链构象多态性方法鉴别研究  

Identification of Clematis Species by PCR-SSCP Analysis

作　　者：安蕊[1] 陈美兰[2] 王学勇[1] 张晓芹[1] 刘春生[1] 

机构地区：[1]北京中医药大学中药学院,北京100102 [2]中国中医研究院中药研究所,北京100700

出　　处：《中国药学杂志》2013年第17期1439-1441,共3页Chinese Pharmaceutical Journal

基　　金：国家科技基础性项目(SB2007FY020)

摘　　要：目的利用聚合酶链反应-单链构象多态性方法(PCR-SSCP)技术建立一种快速、准确鉴别威灵仙药材基原的方法。方法查找Genbank上注册的威灵仙药材基原的内转录间隔区(ITS)序列,进行序列的比对分析,确定稳定的特异鉴别位点。据此设计能扩增包含特异鉴别位点的短片段引物,采用聚合酶链反应-单链构象多态性方法对威灵仙药材基原进行鉴定。结果不同威灵仙基原的聚合酶链反应-单链构象多态性方法谱带带型存在明显差异。结论聚合酶链反应-单链构象多态性方法方法能够快速、准确地鉴别威灵仙药材基原,为进一步开展威灵仙药材的准确鉴定奠定基础。OBJECTIVE To establish a rapid and accurate identification method to distinguish species of Clematis. METHODS First, the registered ITS sequences of Clematis were seeked in the genbank database and compared with those of three species of Clematis＇ to comfirm the stable specific identification site. Second, specific primers were designed, and PCR-SSCP was used to test Clematis. RESULTS The fingerprints of the samples existed significant differences, indicating that our method was able to tell apart the different species of Clematis. CONCLUSION PCR-SSCP has the advantages of high specificity and good reproducibility, there- fore, it lays the foundation for the further development of accurate identification of Clematis.

关 键 词：威灵仙属 东北铁线莲 棉团铁线莲 ITS序列 聚合酶链反应-单链构象多态性方法 聚丙烯酰胺凝胶 分子鉴定 

分 类 号：R28[医药卫生—中药学]