中药材蛇胆的DNA分子标记鉴定研究

摘要

AIM　It is difficult to identify the Chinese crude drug snake gallbladder accurately by morphological and microscopical characteristics or chemical components only. In order to solve the problem, the technique based on DNA molecular marker was introduced into the authentication of snake gallbladder. METHODS　DNA templates were extracted from the membrane or the bile of snake gallbladder, and also from the muscle of the original animal Elaphe schrenckii. About 400 bp DNA fragments of 12S rRNA gene were amplified from the templates and sequenced subsequently. RESULTS　Enough amounts of DNA templates could be extracted from a bit of membrane or bile of snake gallbladder. The sequence of amplicons from the membrane, bile and muscle of the same individual were identical completely. CONCLUSION　The technique of DNA molecular marker could be used for the authentication of snake gallbladder and bile. The results indicate that the technique could be used for the identification of crude drugs from other animal secretion. DNA sequence analysis also demonstrated that the origins of commercial snake gallbladder were complicated and more efficient quality control was necessary for supervising the crude drug in the market.%目的运用分子标记技术鉴定药材蛇胆的真伪，以克服目前仅依据形态、显微特征及理化性状进行鉴别的不足。方法分别从药材蛇胆的胆衣和胆汁、原动物棕黑锦蛇的肌肉和胆汁中提取DNA，经PCR扩增得到约400 bp的12S rRNA基因片段，并对该基因片段进行测序研究。结果从少量药材蛇胆的胆衣和胆汁中可提得足够用于PCR扩增的DNA模板，扩增产物的测序结果表明同一动物的胆衣和胆汁、肌肉和胆汁的碱基序列完全一致。结论 DNA分子标记技术可用于中药材蛇胆和胆汁的鉴定，提示该技术也可用于其他动物分泌物类型药材的鉴别。目前市场上药材蛇胆来源较复杂，需加强质量监督和控制。