摘要
目的探讨鹿鞭及牛鞭线粒体细胞色素b基因部分序列片段特征,建立中药材鹿鞭敏感、特异的DNA分子标记学鉴定方法。方法碱变性法提取新鲜鹿鞭及牛鞭,采用引物设计软件PrimerPremier5.0对鹿鞭及牛鞭的细胞色素b基因序列分析,用于鉴定正品鹿鞭的特异性引物,经聚合酶链式反应(PCR)扩增,对鹿鞭进行DNA指纹鉴定。结果对鹿鞭进行mtDNA鉴定图谱显示,真品鹿鞭有306bp条带,伪品没有。结论用此方法对鹿鞭进行鉴定,可准确辨别正品与伪品。重现性、稳定性好,简便准确而可行。鹿鞭mtDNA具有特异性指纹特征,所得鹿鞭DNA指纹特征图谱可用于鹿鞭的鉴定。
Objective To explore the identification and characterization of cytochrome gene from the testis et penis cervi and to develop a method of DNA molecular marker for identification of traditional chinese medicine,the testis et penis cervi.Methods A pair of allele-specific primers was designed for distinguishing the genuine drugs from their adulterants with primer design ware premier 5.0.The fresh testis et penis cervi and the bullwhip were extracted and polymerase chain reaction（PCR）amplification was carried out.The DNA bands of testis et penis cervi were recovered,purified and sequenced.Commercially available six samples were identificated.Results The results of diagnostic PCR for original animals showed that a 306 bp fragment was only amplified from DNA templates of the testis et penis cervi Only 3 samples were genuine drug.Conclusion The method were simple and the result is reliable.It could be used for the crude drugs in the current market.
出处
《中国医药科学》
2011年第19期112-113,共2页
China Medicine And Pharmacy
基金
吉林省科技发展计划项目(20090906)
关键词
鹿鞭
线粒体DNA
鉴别引物
PCR
DNA指纹鉴定
Testis et penis cervi
MtDNA
Diagnostic primers
PCR
DNA fingerprinting identification
