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

Abstract

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

CLC Number:

S826.2

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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Establishment of the Efficient Method to Detect the FecB Mutation and the Construction of the Prolific Tan Sheep Population

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