基于SSR标记技术对3种国产绵羊肉的品种鉴别分析  

Identification of Ovine Livers from Three Indigenous Chinese Breeds Based on Simple Sequence Repeat Markers

作　　者：宋亚倩 杨波[1] 罗瑞明[1] 马小梅[1] SONG Yaqian;YANG Bo;LUO Ruiming;MA Xiaomei(School of Food and Wine,Ningxia University,Yinchuan 750021,China)

机构地区：[1]宁夏大学食品与葡萄酒学院,宁夏银川750021

出　　处：《食品科学》2022年第4期346-354,共9页Food Science

基　　金：宁夏回族自治区创新平台专项(2019DPC05011);宁夏肉品加工及质量安全控制工程技术研究中心科研项目(2019DPC05011)。

摘　　要：提取纯种宁夏滩羊、纯种新疆细毛羊和纯种藏绵羊的肝脏DNA,并进行文库构建、扩增、纯化和质控。检测序列中的简单重复序列(simple sequence repeats,SSR)位点,设计合成并筛选50对引物进行聚合酶链式反应扩增并使用毛细管电泳检测SSR分型后,挑选多态性引物对宁夏滩羊、新疆细毛羊和藏绵羊样本进行遗传多态性检测。结果显示:有10对SSR引物成功扩增出稳定条带,分析10对多态性SSR引物的遗传多样性得出,宁夏滩羊、新疆细毛羊和藏绵羊的平均等位基因数分别为4.100、3.100和3.600;平均有效等位基因数分别为3.782、2.747和3.193;平均观测杂合度分别为1.007、1.009和0.953;平均期望杂合度分别为0.704、0.615和0.660;平均多态信息含量分别为0.432、0.414和0.425,说明宁夏滩羊的遗传多态程度高于藏绵羊和新疆细毛羊。毛细管荧光电泳检测峰图结果表明:scaffold632:150-463荧光引物对宁夏滩羊源性肉特异性良好;scaffold31384:146-461荧光引物对新疆细毛羊源性肉特异性良好;scaffold7676:103-270荧光引物对藏绵羊肉特异性良好,这3对引物能够实现对宁夏滩羊、新疆细毛羊和藏绵羊的有效区分。建立的我国西北地区3个绵羊品种基因库,可以用于科学准确判断3个纯种绵羊肉品种。In this study, liver DNA from purebred Ningxia Tan sheep, purebred Xinjiang fine-wool sheep and purebred Tibetan sheep was extracted followed by genomic library construction, amplification, purification and quality control.Simple sequence repeat(SSR) sites in the sequence were detected, and 50 pairs of primers were designed, synthesized and screened for polymerase chain reaction(PCR) amplification. SSR typing was carried out by capillary electrophoresis, and polymorphic primers were selected to detect the genetic polymorphism of the three breeds. The results showed that 10 pairs of SSR primers successfully amplified stable bands. Ten pairs of polymorphic SSR primers were used to analyze the genetic diversity of Ningxia Tan sheep, Xinjiang fine-wool sheep and Tibetan sheep, showing an average number of alleles(Na) of 4.100, 3.100 and 3.600, average number of effective alleles(Ne) of 3.782, 2.747 and 3.193, average observed heterozygosity(Ho) of 1.007, 1.009 and 0.953, average expected heterozygosity(He) of 0.704, 0.615 and 0.660, and average polymorphic information content(PIC) of 0.432, 0.414 and 0.425, respectively. These data indicated that the genetic polymorphism of Ningxia Tan sheep was higher than that of Tibetan sheep and Xinjiang fine-wool sheep. The results of capillary electrophoresis with fluorescence detection showed that the fluorescent primers scaffold632:150-463, scaffold31384:146-461 and scaffold7676:103-270 had excellent specificity for Ningxia Tan, Xinjiang fine-wool and Tibetan sheep, respectively, and they could effectively distinguish the breeds. In conclusion, the gene banks established can allow scientific and accurate identification of Ningxia Tan sheep, Tibetan sheep and Xinjiang Fine-wool sheep.

关 键 词：基因组DNA 微卫星序列 特异性检测 绵羊肉品种鉴别 

分 类 号：TS251.5[轻工技术与工程—农产品加工及贮藏工程]