Rapid Identification of the SNP Mutation in the ABCD4 Gene and Its Association with Multi-Vertebrae Phenotypes in Ujimqin Sheep Using TaqMan-MGB Technology

文献类型： 外文期刊

第一作者： Zhang, Yue

作者： Zhang, Yue;Liu, Jun;Zhang, Yong;Zhang, Yue;Zhang, Min;Su, Hong;Zhao, Feifei;Zhao, Yifan;Li, Xiunan;Cao, Guifang;Zhang, Yue;Zhang, Min;Su, Hong;Zhao, Feifei;Zhao, Yifan;Li, Xiunan;Cao, Guifang;Li, Xiunan;Yang, Yanyan;Cao, Guifang;Zhang, Yong

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关键词： Ujimqin sheep; multi-vertebrae; ABCD4 gene; single nucleotide polymorphism; TaqMan-MGB detection; molecular marker

期刊名称：ANIMALS （ 影响因子：2.7； 五年影响因子：3.2 ）

ISSN： 2076-2615

年卷期： 2025 年 15 卷 15 期

页码：

收录情况： SCI

摘要： Simple Summary This study developed a fast, cost-effective genetic test to identify Ujimqin sheep with extra vertebrae, a trait linked to higher meat yield. Researchers targeted a specific gene change (SNP: ABCD4, Chr7:89393414 C > T) known to cause extra vertebrae. We designed a TaqMan-MGB test using fluorescent probes to detect this SNP in blood DNA. Validated on 152 sheep, the test showed 100% agreement with gold-standard DNA sequencing (Sanger method). Compared to DR imaging (83.6% overall agreement), it accurately detected 91.4% of multi-vertebrae sheep (127/139 animals). The test is simple, high-throughput, and significantly cheaper than DR imaging. This tool enables early, efficient selection of breeding sheep with the desirable multi-vertebrae trait, accelerating genetic improvement programs for Ujimqin sheep farmers. Ujimqin sheep, known for its distinctive multi-vertebrae phenotypes (T13L7, T14L6, and T14L7) and economic value, has garnered significant attention. However, conventional phenotypic detection methods suffer from low efficiency and high costs. In this study, based on a key SNP locus (ABCD4 gene, Chr7:89393414, C > T) identified through a genome-wide association study (GWAS), a TaqMan-MGB (minor groove binder) genotyping system was developed. the objective was to establish a high-throughput and efficient molecular marker-assisted selection (MAS) tool. Specific primers and dual fluorescent probes were designed to optimize the reaction system. Standard plasmids were adopted to validate genotyping accuracy. A total of 152 Ujimqin sheep were subjected to TaqMan-MGB genotyping, digital radiography (DR) imaging, and Sanger sequencing. the results showed complete concordance between TaqMan-MGB and Sanger sequencing, with an overall agreement rate of 83.6% with DR imaging. For individuals with T/T genotypes (127/139), the detection accuracy reached 91.4%. This method demonstrated high specificity, simplicity, and cost-efficiency, significantly reducing the time and financial burden associated with traditional imaging-based approaches. the findings indicate that the TaqMan-MGB technique can accurately identify the T/T genotype at the SNP site and its strong association with the multi-vertebrae phenotypes, offering an effective and reliable tool for molecular breeding of Ujimqin sheep.

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