林蛙油的DNA提取及鉴定方法

DNA extraction and identification of the oviductus ranae

中文关键词:  东北林蛙  林蛙油  线粒体COI基因  DNA测序  真伪鉴别

英文关键词:Rana dybowskii  oviductus ranae  Mitochondrial COI gene  DNA sequencing  authenticity identification

中文摘要:

      目的 建立基于分子生物学鉴定林蛙油真伪的检测方法。方法 采集成年雌蛙卵巢组织, 经风干制成林蛙油, 通过改良方法提取总DNA, 以已报道的东北林蛙、黑龙江林蛙、黑斑侧褶蛙、牛蛙和中华蟾蜍的COI及cytB基因为模板, 设计引物探针。结果 PCR扩增产物测序结果中仅东北林蛙获得230 bp左右产物, 且序列比对与东北林蛙COI序列一致性为99%左右, 说明实验对象林蛙为东北林蛙。结论 利用线粒体基因独特的优势, 应用PCR扩增东北林蛙COI基因, 通过测序确定东北林蛙亚种, 建立了东北林蛙亚种及其产品的鉴别方法。

英文摘要:

      Objective To establish a detection method based on molecular biology to identify the authenticity of oviductus ranae. Methods The ovary tissues of adult female frogs were collected and dried to make frog oil. The total DNA was extracted by the improved method. Primer probes were designed using the reported COI and cytB genes of Rana dybowskii, Rana amurensis, Pelophylax nigromaculatus, Rana (Lithobates) catesbeianus and Bufo raddei as templates. Results Sequencing results of PCR amplification products showed that only Rana dybowskii obtained about 230 bp of products, and the sequence alignment was about 99% consistent with the COI sequence of Rana dybowskii, indicating that the experimental object was Rana dybowskii. Conclusion Based on the unique advantages of mitochondrial gene, the COI gene of Rana dybowskii is amplified by PCR, and the subspecies of Rana dybowskii is identified by sequencing. The identification method of Rana dybowskii and its products is established.