哈蟆油基原物种DNA分子鉴定研究  

DNA molecular identification of Ranae oviductus original species

作　　者：高晓晨[1] 刘冬[1] 白俊武[2] 于江波[2] 赵娜娜[1] 宗颖[1] 崔丽娜[1] 张辉[1] 孙佳明[1] 

机构地区：[1]长春中医药大学,长春130117 [2]吉林敖东延边药业股份有限公司,吉林敦化133700

出　　处：《吉林中医药》2014年第11期1147-1148,1162,共3页Jilin Journal of Chinese Medicine

基　　金：吉林省中医药管理局中医药科技项目:蛤蟆油原动物及其混为品的COI序列分子鉴定(YJS-0228);国家中医药管理局行业专项:<中国药典>名贵珍稀动物药及混伪品鉴定技术规范应用研究(201407007)

摘　　要：目的建立一种简单、可行的蛤蟆油基原物种DNA分子鉴定方法,并为其鉴定提供有效的手段。方法以COI作为条形码序列,对蛤蟆油基原物种中国林蛙和黑龙江林蛙进行DNA提取,PCR扩增和双向测序,用CodonCode Aligner V 4.2.7校对拼接,并用MEGA6.0对其样品进行种内分析。结果实验样品可以获得蛤蟆油基原物种的序列,中国林蛙有34个碱基变异位点,种内平均K2P距离为0.042,种内最大K2P距离为0.058。黑龙江林蛙有3个碱基变异位点,种内平均K2P距离为0.003,种内最大K2P距离为0.005。结论运用COI条形码序列能够准确鉴定蛤蟆油基原物种,为其鉴定提供新方法。Objective To establish a simple and feasible method of DNA molecular identification to identify the Ranae oviductus original species and provide effective measures for its identification. Methods With COI as the barcode sequence,DNA was extracted from the experimental samples（ Rana temporaria chensinensis and Rana amurensis Boulenger）. The COI sequences were amplified and sequenced bi-directionally. Sequence alignment and NJ tree construction were carried out by MEGA6. 0 software. Results COI sequences can be obtained from all experimental samples.Rana temporaria chensinensis David has 34 base mutation loci. The average intra-specific K2 P distance was 0. 042 and the maximum intra-specific distance was 0. 058. Rana amurensis Boulenger has 3 base mutation loci. The average intra-specific K2 P distance was 0. 003 and the maximum intra-specific distance was 0. 005. Conclusion COI can be used correctly to identify Ranae oviductus original species and provide new method for its identification.

关 键 词：哈蟆油 基原物种 DNA鉴定 碱基 中国林蛙 黑龙江林蛙 

分 类 号：R284.1[医药卫生—中药学]