基于线粒体12SrRNA基因鉴别混合牛肉与制品的牛种来源.pdfVIP

HEREDITAS (Beijing) 2008 8 , 30(8): 1008―1014 ISSN 0253-9772 研究报告 DOI: 10.3724/SP.J.1005.2008.01008 12S rRNA 1 2 1 1 1 陈冬, 柏凡 , 周明亮 , 张翔宇, 吴登俊 1. , 625014; 2. , 610041 : 线粒体基因组具有种内高度保守性。它的 12S rRNA 基因同时具有抗腐蚀、耐高温的特点, 而被用于饲 料、鲜肉及肉制品的来源追踪和物种成分鉴别等研究。利用牦牛、普通牛、水牛作为研究材料, 在牛线粒体 12S rRNA 基因通用引物扩增片段区域发现 3 种特异的酶切位点, 可用于混合鲜牛肉及制品的牛种来源的鉴别。结 果显示: 牦牛12S rRNA 扩增片段被酶切为203 bp 和250 bp, 普通牛为134 bp 和318 bp, 水牛为86 bp 和367 bp, 通过测序验证了特异性位点和酶切的正确性; 对样品经过不同温度(100 、120 、140 、160 和 180 )处 理均能扩增到目的条带, 但条带从 120 开始变淡。该方法在鲜牛肉及制品的牛种来源鉴别上具有简单、快速、 廉价等优点。 : 12S rRNA; 酶切; 牛线粒体基因; 内源参照基因 Differentiation of Bos grunniens, Bos Taurus, and Bubalus from meat products mixture based on mitochondrion 12S rRNA gene 1 2 1 1 1 CHEN Dong , BAI Fan , ZHOU Ming-Liang , ZHANG Xiang-Yu , WU Deng-Jun 1. College of Animal Science and Technology, Sichuan Agricultural University, Ya ’an 625014, China; 2. Sichuan Feedstuff Workstation, Chengdu 610041, China Abstract: Mitochondrial DNA sequence is highly conserved within species. Gene 12S rRNA is able to endure degeneration and high temperature, which allows identification of feedstuff, fresh meat, processed meat, and traceability. In the present study, three unique restriction sites were detected in the fragments of mitochondrial 12S rRNA gene regions amplified with universal primer, which were able to distinguish Bos grunniens, Bos. taurus, and Bubalus in fresh meat and processed meat mixture. The fragment of yak was digested to 134 bp and 318 bp, scalper 134 bp and 318 bp, and buffalo 86 bp and 367 bp. The specific locus and digestion