Differentiation of European wild boar (Sus scrofa scrofa) and domestic swine (Sus scrofa domestica) meats by PCR analysis targeting the mitochondrial D-loop and the nuclear melanocortin receptor 1 (MC1R) genes
Differentiation of European wild boar (Sus scrofa scrofa) and domestic swine (Sus scrofa domestica) meats by PCR analysis targeting the mitochondrial D-loop and the nuclear melanocortin receptor 1 (MC1R) genes
复制标题
DOI：
10.1016/j.meatsci.2007.06.018
复制
发表时间：
2008-03-01
期刊：
影响因子：
7.1
通讯作者：
Martin, Rosario
中科院分区：
文献类型：
--
作者：
Fajardo, Violeta;Gonzalez, Isabel;Martin, Rosario
本研究利用线粒体位移环（D-loop）区和核黑素皮质素受体1 （MC1R）基因两个分子标记的PCR靶向序列，描述了欧洲野猪（Sus scrofa scrofa）和家猪（Sus scrofa domestica）肉的分化。我们在野生和家养的苏氏丝瓜肉样品中扩增了一个多态d环片段（约270 bp），并对其进行了测序，以寻找适合PCR-RFLP分析的核苷酸区域。序列数据显示，在Sus scrofa D-loop序列中仅存在少量点突变，无法直接区分野猪肉和家猪肉。随后，以MC1R基因为靶点，构建scro特异性引物，扩增795 by MC1R片段。随后对MO R猪特异性扩增子的RFLP分析允许选择BspHI和BstUI内切酶进行种内Sus scrofa分化。用所选择的酶消化MC1R扩增子产生特异性PCR-RFLP图谱，可以区分野生猪和家猪标本的肉类。该技术还能够检测出产生杂合谱的样本，这表明野猪和家猪杂交产生。本文报道的针对MO R基因的PCR-RFLP可以常规地用于验证游戏产品的正确标签。(c) 2007 Elsevier Ltd.版权所有。
This work describes the differentiation of European wild boar (Sus scrofa scrofa) and domestic swine (Sus scrofa domestica) meats by PCR targeting sequences from two molecular markers: the mitochondrial displacement loop (D-loop) region and the nuclear melanocortin receptor 1 (MC1R) gene. A polymorphic D-loop fragment (similar to 270 bp) was amplified and sequenced in a number of wild and domestic Sus scrofa meat samples, to find a nucleotide region suitable for PCR-RFLP analysis. Sequence data showed the presence of only a few point mutations across Sus scrofa D-loop sequences, not allowing direct discrimination between wild boar and domestic swine meats. Later, the MC1R gene was targeted and Sits scrofa-specific primers designed to amplify a 795 by MC1R fragment. Subsequent RFLP analysis of the MO R swine-specific amplicons allowed selection of BspHI and BstUI endonucleases to carry out intraspecific Sus scrofa differentiation. Digestion of MC1R amplicons with the chosen enzymes generated characteristic PCR-RFLP profiles that allowed discrimination among meats from wild and domestic swine specimens. The technique also enabled the detection of samples that yielded heterozygous profiles, suggesting hybrids resulting from wild boar and domestic pig breeding. The PCR-RFLP reported here, targeting the MO R gene may be routinely applied to verify the correct labelling of game products. (c) 2007 Elsevier Ltd. All rights reserved.