林蛙油的DNA提取及鉴定方法  

DNA extraction and identification of the oviductus ranae

作　　者：聂丹丹 王春雨 毕程程 胡婷婷 付海滨 李玲 NIE Dan-Dan;WANG Chun-Yu;BI Cheng-Cheng;HU Ting-Ting;FU Hai-Bin;LI Ling(Changchun Customs Technical Center,Changchun 130062,China;Changchun Customs District People’s Republic of China,Changchun 130033,China;Yanji customs comprehensive technical services,Yanji 133001,China;Shenyang Customs Technical Center,Shenyang 110000,China)

机构地区：[1]长春海关技术中心,长春130062 [2]长春海关,长春130033 [3]延吉海关技术中心,延吉133001 [4]沈阳海关技术中心,沈阳110000

出　　处：《食品安全质量检测学报》2021年第5期1936-1944,共9页Journal of Food Safety and Quality

基　　金：原国家质检总局项目(2017IK208)。

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。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.

关 键 词：东北林蛙 林蛙油 线粒体COI基因 DNA测序 真伪鉴别 

分 类 号：R282.5[医药卫生—中药学] TS225.2[医药卫生—中医学]