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作 者:生书晶[1,2] 严萍[1] 郑传进[1] 赵树进[1]
机构地区:[1]中国人民解放军广州军区广州总医院,广东广州510010 [2]华南理工大学生物科学与工程学院,广东广州510641
出 处:《中药材》2010年第11期1707-1711,共5页Journal of Chinese Medicinal Materials
基 金:广东省自然科学基金项目(06019716)
摘 要:目的:比较何首乌与其常见混淆品的matK基因序列,从分子水平对中药材何首乌进行鉴定。方法:改良的CTAB法提取基因组DNA,用一对matK通用引物进行PCR扩增,TA克隆后进行测序和序列分析。结果:除毛脉蓼之外,其他混淆品与正品何首乌间的matK基因序列差异较大,不论是核苷酸替代数还是遗传距离,都远远大于不同居群的何首乌之间;进一步比对分析,发现了可以快速鉴别何首乌及其同属混淆品的鉴别位点;运用matK序列分析成功地对3种市售何首乌药材进行了真伪鉴定。结论:根据matK序列特征,能有效区分何首乌及其常见混淆品,matK序列可以作为何首乌药材鉴定的分子标记。Objective:To identify Fallopia multiflora from its adulterants by comparing their matK sequences.Methods: Genomic DNA of different materials was extracted using modified cetyltrimethyl ammonium bromide(CTAB) method.The double-strand matK genes were amplified using PCR method and then sequenced.The data were analyzed in Clustral W and MEGA 4.0 software package.Results: Besides F.multiflora var.ciliinerve,the matK sequences of other adulterants show distinct differences with F.multiflora,whether for nucleotides substitutions or genetic distances;and the specific identifying sites for distinguishing F.multiflora and other Fallopia adulterants were found through further comparative analysis.Moreover,the 3 inspected materials were successfully authenticated by comparing the matK sequences.Conclusion:matK sequences can be used for the molecular identification between F.multiflora and its adulterant species.
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