地龙类药用动物的简单序列重复区间分子鉴定研究  

Identification of Medicinal Animals of Pheretima by Inter-Simple Sequence Repeat Markers

作　　者：吴文如[1] 李薇[1] 赖小平[1] 

机构地区：[1]广州中医药大学中药学院,广东广州510006

出　　处：《广州中医药大学学报》2011年第4期423-426,共4页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(编号:30772741);国家科技部支撑计划项目(编号:2006BAI061102);广东省科技计划项目(编号:2007B020701004)

摘　　要：【目的】探索利用简单序列重复区间(ISSR)分子标记方法在核酸分子水平上鉴别不同来源的地龙类药用动物。【方法】从100条ISSR引物中筛选出能扩增明显多态性条带的引物,对采集自广东、福建、湖北、上海等地的7个不同品种的地龙类药用动物样品进行扩增和凝胶电泳分析,寻找特征位点。采用NTSYS软件计算材料间的相似系数,并用UPGMA法构建系统树,按遗传距离构建分子聚类图。【结果】8条引物共扩增出137个DNA片段,其中127个片段呈现多态性,平均多态位点百分率为92.35%,可单独或联合应用鉴别地龙类药用动物。【结论】ISSR分子标记技术具有较好的可靠性和重现性,可用于地龙类药用动物的鉴别和遗传亲缘关系研究。Objective To identify medicinal animals of Pheretima by inter-simple sequence repeat（ISSR） markers at nucleinic acid molecular level.Methods Primers suitable for routine analysis were screened from 100 inter-simple sequence repeat primers.Then the selected primers were used for PCR and agarose electrophoresis of 7 kinds of medicinal animals of Pheretima which were collected mainly from Guangdong,Fujian,and Hubei.The systematic diagram of similarity coefficient was set up by NTSYS software and genetic distance was clustered by UPGMA method.Results The amplification of 8 ISSR primers screened out of 100 primers resulted in 137 electrophoresis strips with 127 strips（92.35%） polymorphic.Conclusion Inter-simple sequence repeat markers provide quick and reliable molecular markers for identification of medicinal animals of Pheretima.

关 键 词：地龙 简单序列重复区间 分子鉴定 基因扩增 

分 类 号：R282.5[医药卫生—中药学]