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

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.