鹿茸线粒体DNA的指纹鉴定研究  

Identification of Velvet Antler by Mitochondrial DNA Fingerprint

作　　者：谷玉娟[1] 张丽华[1] 傅桂莲[2] 

机构地区：[1]北华大学药学院,吉林吉林132013 [2]北华大学检验学院,吉林吉林132013

出　　处：《中国药学杂志》2013年第3期170-173,共4页Chinese Pharmaceutical Journal

基　　金：吉林省科技发展计划项目(20090906)

摘　　要：目的通过鹿茸特异性引物鉴别和随机扩增多态DNA(RAPD)鉴别的比较,选择一种更简便的方法用于鉴别鹿茸。方法采用盐析法从梅花鹿茸、马鹿茸、驯鹿茸、市售鹿茸等样品中抽提线粒体DNA,并应用试剂盒进行纯化。进行特异性引物扩增和序列测定,同时进行随机扩增多态DNA扩增。结果梅花鹿茸,马鹿茸,驯鹿茸以及部分市售鹿茸经特异性引物扩增后均可在琼脂糖凝胶中显示313 bp片段,只是条带亮度不同。而其余市售鹿茸无扩增条带;随机扩增多态DNA不仅有效显示阳性与阴性的DNA扩增结果,而且对梅花鹿茸,马鹿茸,驯鹿茸的聚合酶链反应产物的差异,以不同的条带数目和条带亮度得以验证。结论随机扩增多态DNA鉴别鹿茸真伪的方法更准确快捷,通过随机扩增多态DNA扩增后主条带与梅花鹿茸或马鹿茸扩增的条带大小和亮度一致,则为正品;如不一致,则为《中国药典》规定外的鹿茸或其他混淆品。这种方法对于筛选、甄别市售动物中药材,特别是名贵中药具有广阔的应用前景。OBJECTIVE To compare specific primer method and random amplified polymorphic DNA （RAPD） method for identification of Velvet Antler, thus to choose a simpler method. METHODS DNA samples were extracted from sika deer antler, wild deer antler, reideer pilose antler and commercially available Velvet Antler, and then purified and amplified with specific primer ampli- fication and RAPD amplification. RESULTS The sika deer antler, wild deer antler, reideer pilose antler and some of the commercially available Velvet Antler all showed 313 bp segment in the agarose gel with different brightness while the other commercial products did not show this segment. RAPD not only effectively showed positive and negative DNA amplification results, but also showed the difference in PCR products of sika deer antlerwild deer antler with different strip numbers and strip brightness. CONCLUSION RAPD method is more accurate and rapid and it has wide application prospect. KEY WORDS： Velvet Antler; specific primer; random amplified polymorphic DNA（RAPD） ; PCR

关 键 词：鹿茸 特异性引物 随机扩增多态DNA 聚合酶链反应 

分 类 号：R284[医药卫生—中药学]