绵羊FecB突变检测方法的建立及高繁滩羊核心群的构建

doi:10.11843/j.issn.0366-6964.2022.09.010

摘要/Abstract

摘要： 旨在建立快速、高效、精准检测FecB突变的荧光qPCR技术，为滩羊多羔性能研究、群体改良及肉用多羔滩羊新品系培育提供技术支撑。基于荧光qPCR技术基本原理，设计特异性引物对，开发基于荧光qPCR检测绵羊FecB突变的方法；对85只已知FecB基因型的健康、适繁雌性滩羊血液基因组DNA样品分别采用PCR-Sanger测序法、TaqMan探针法和荧光qPCR法进行FecB基因分型，统计3种方法的准确率；采用开发的荧光qPCR方法对939只健康、适繁滩羊进行FecB基因分型，统计不同FecB基因型经产母羊的产羔率。结果表明，使用TaqMan探针法、PCR-Sanger测序法和开发的荧光qPCR法对已知FecB基因型样品检测的比较中，荧光qPCR法对各基因型鉴定的准确度均达100%，高于前两者。939只滩羊FecB基因分型结果显示，6只滩羊为FecB突变纯合型（BB）、57只为杂合型（B+）、876只为野生型（++），BB型、B+型和++型滩羊的基因型频率分别为0.64%、6.07%和93.29%，其中B等位基因频率为0.04，+等位基因频率为0.96；监测母羊群体的产羔数发现，FecB突变纯合型滩羊产羔率为166.67%，突变杂合型滩羊产羔率为153.12%，野生型滩羊产羔率为105.78%，纯合突变型和杂合突变型母羊群体的产羔率极显著高于野生型群体（P<0.01）。综上所述，与TaqMan探针法及PCR-Sanger测序法相比，本研究建立的绵羊FecB突变荧光qPCR分型方法具有简便易行、廉价高效的优点，在绵羊的分子选育中具有较高应用价值；同时本研究证实了滩羊FecB突变可显著提高母羊产羔率，为多羔滩羊的分子选育提供了坚实的技术支撑。

关键词: FecB, 荧光qPCR, 特异引物, 滩羊

Abstract: The aim of the current study was to establish a fluorescent qPCR technology for rapid, efficient and precise detection of the FecB mutation. This will provide technical support for Tan sheep multi-lamb performance research, population improvement and breeding of improved prolific and meat-producing Tan sheep strains. In this study, a method for detecting the FecB mutation in Tan sheep based on fluorescence qPCR was developed by designing specific primer pairs. Genomic DNA samples of 85 healthy and reproducible Tan ewes with known FecB genotypes were genotyped by PCR-based Sanger sequencing, TaqMan probe and fluorescence qPCR, respectively. Subsequently, the accuracy of the 3 methods was detected. Additionally, the FecB genotyping of 939 Tan sheep was carried out by the developed fluorescent qPCR method. Furthermore, the lambing rates of primiparous ewes with different FecB genotypes were recorded. TaqMan probe, PCR-based Sanger sequencing and the developed fluorescence qPCR method were used to detect the FecB mutation in samples with known genotypes. The results showed that the accuracy of the developed fluorescent qPCR method for the identification of each FecB genotype was 100%, which was higher than the other two methods. The results of genotyping the FecB mutation in 939 healthy and reproducible Tan ewes showed that 6 ewes were homozygous (BB), 57 ewes were heterozygous (B+), and 876 ewes were wild type (++). The genotype frequencies of BB, B+ and ++ Tan sheep were 0.64%, 6.07% and 93.29%, respectively, of which the B allele frequency was 0.04 and the + allele frequency was 0.96. The lambing of the ewes was monitored, it was found that the lambing rate of Tan sheep with FecB mutant homozygous was 166.67%, the lambing rate of Tan sheep with mutant heterozygous was 153.12%, and the lambing rate of wild-type Tan sheep was 105.78%. The lambing rate of the ewes with mutant homozygous and heterozygous was significantly higher than that of the wild-type Tan sheep (P<0.01).Compared with TaqMan and PCR-based Sanger sequencing methods, the developed fluorescence qPCR method in this study has the advantages of simplicity, low cost, and effectiveness. Thus, it has potential application value for molecular breeding. Therefore, the established fluorescent qPCR detection method provides a solid technical support for detecting the FecB genotypes in sheep. Furthermore, it is confirmed that the mutation of FecB can significantly improve the lambing rate of local sheep breeds.

Key words: FecB mutation, fluorescent qPCR, specific primers, Tan sheep

中图分类号:

S826.2

周世卫, 吴庭杰, 王倩, 蔡蓓, 韩赛铮, 吴彦虎, 王小龙, 陈玉林. 绵羊FecB突变检测方法的建立及高繁滩羊核心群的构建[J]. 畜牧兽医学报, 2022, 53(9): 2920-2929.

ZHOU Shiwei, WU Tingjie, WANG Qian, CAI Bei, HAN Saizheng, WU Yanhu, WANG Xiaolong, CHEN Yulin. Establishment of the Efficient Method to Detect the FecB Mutation and the Construction of the Prolific Tan Sheep Population[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 2920-2929.

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