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基于线粒体 COI 基因的 PCR-RFLP 技术鉴定肉类物种。

Identification of meat species by PCR-RFLP of the mitochondrial COI gene.

机构信息

Department of Molecular Biology and Biotechnology, AECS, PO Box 6091, Damascus, Syria.

出版信息

Meat Sci. 2012 Feb;90(2):490-3. doi: 10.1016/j.meatsci.2011.09.013. Epub 2011 Sep 22.

Abstract

Meat authenticity verification is pertinent for economical, religious or public health concerns. The present study investigates the use of PCR-RFLP of a part of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene for identification of species origin of raw meat samples of cow, chicken, turkey, sheep, pig, buffalo, camel and donkey. PCR yielded a 710-bp fragment in all species. The amplicons were digested with seven restriction endonucleases (Hind II, Ava II, Rsa I, Taq I, Hpa II, Tru 1I and Xba I) that were selected based on the preliminary in silico analysis. Different levels of polymorphism were detected among samples. The level of COI variation revealed using only Hpa II was sufficient to generate easily analyzable species-specific restriction profiles that could distinguish unambiguously all targeted species. Compared to previously published reports for the determination of meat origin at the molecular level, the approach developed here is much cheaper and faster for routine identification of meats in food control laboratories.

摘要

肉类真实性验证与经济、宗教或公共卫生问题息息相关。本研究利用线粒体细胞色素 c 氧化酶亚基 1(COI)基因的部分 PCR-RFLP 技术,鉴定了牛、鸡、火鸡、羊、猪、水牛、骆驼和驴等生肉样品的物种来源。PCR 在所有物种中均产生了 710bp 的片段。根据初步的计算机分析,选择了七种限制性内切酶(Hind II、Ava II、Rsa I、Taq I、Hpa II、Tru 1I 和 Xba I)对扩增子进行酶切。在样本中检测到不同水平的多态性。仅使用 Hpa II 检测到的 COI 变异水平足以生成易于分析的种特异性限制性图谱,可明确区分所有目标物种。与之前在分子水平上用于确定肉类来源的已发表报告相比,该方法在食品控制实验室中用于常规肉类鉴定时更便宜、更快。

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