基于COⅠ基因的羚羊角及其混伪品的DNA条形码鉴定方法的研究
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篇名: 基于COⅠ基因的羚羊角及其混伪品的DNA条形码鉴定方法的研究
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摘要: 目的:考察利用细胞色素C氧化酶亚基Ⅰ(COⅠ)基因对羚羊角及其混伪品进行分子鉴定的可行性。方法:收集4个地区的羊角类整角、不完整角样品,共7个。比较样品骨塞部位和角质层部位DNA的提取效果,采用聚合酶链式反应(PCR)技术,以通用引物LCOⅠ490、HCO2198扩增样品的COⅠ基因,通过凝胶电泳、纯化(取750 bp条带)、测序后,以COⅠ基因作为条形码比对序列,采用NCBI数据库中的Blast软件在线比对,确定收集样品的具体物种。结果:样品以角质层部位进行DNA提取的效果较好(DNA浓度为15.7~22.6 ng/μL,260 nm/280 nm吸光度值为1.73~4.72);在线比对结果显示,所有样品COⅠ基因的相似性均达到99%,其主要来源于赛加羚羊、藏羚羊、普氏原羚、山羊、绵羊等羊类的角。结论:基于COⅠ基因的DNA条形码技术可用于羚羊角及其混伪品的鉴定,该技术为羊角类药材的鉴定提供了一种准确、客观的方法。
ABSTRACT: OBJECTIVE: To investigate the feasibility of molecule identication of Saiga tatarica and its adulterants by using cytochrome C oxidase subunit Ⅰ (COⅠ) gene. METHODS:A total of 7 horns and incomplete horns were collected from 4 areas.The extraction effect of DNA from bone plug and stratum corneum were investigated; PCR technology was used to amplify COⅠ gene of samples using universal primer LCOⅠ490, HCO2198; after gel electrophoresis, purification (750 bp strip) and sequencing, using COⅠ gene as barcode alignment sequence, online comparison was conducted by using Blast software of NCBI database to determine specific species. RESULTS:The extraction of DNA from stratum corneum was better (DNA concentration was 15.7-22.6  ng/μL, the absorbance of 260 nm/280 nm was 1.73-4.72). Online comparison showed that the similarity of COⅠ gene in all samples reached 99%, mainly from the horns of saiga antelope, Tibetan antelope, gazelle, sheep and goats. CONCLUSIONS: DNA barcode technology based on COⅠ gene can be used for the identification of S. tatarica and its adulterants. The technology can provide an accurate and objective method for the identification of horn medicinal materials.
期刊: 2018年第29卷第1期
作者: 王桂梅,王世伟,席啸虎,张小慧
AUTHORS: WANG Guimei,WANG Shiwei,XI Xiaohu,ZHANG Xiaohui
关键字: 细胞色素C氧化酶亚基Ⅰ基因;羚羊角;混伪品;序列比对;鉴定
KEYWORDS: Cytochrome C oxidase subunit Ⅰ; Saiga tatarica; Adulterants; Sequence alignment; Identification
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