开发和验证用于确定与绵羊体重相关的 DGKH 和 PPP1R1C 基因多态性的测试系统

O. Koshkina, T. Deniskova, N. Zinovieva
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引用次数: 0

摘要

相关性。考虑到现代 DNA 技术的使用,国家绵羊育种落后于其他畜牧业分支。寻找新基因--与重要经济性状相关的潜在候选基因--对于更全面地揭示绵羊的遗传潜力具有重要意义。早先的一项全基因组关联研究表明,DGKH 和 PPP1R1C 基因对绵羊出生时和 90 日龄时的体重有一定影响。因此,对 DGKH 和 PPP1R1C 基因的多态性进行更详细的研究可以加深对家羊生长发育过程的了解,因此我们选择这两个基因作为实验的目标基因。根据 NCBI 中绵羊第 10 号染色体(NC_056063.1)和第 2 号染色体(NC_056055.1)的参考 DNA 序列,选择引物和探针扩增长度为 68 个核苷酸对的 DGKH 和 PPP1R1C 基因目标 SNP 片段。为确定多态性,开发了基于实时 PCR 的测试系统。基因型通过多参数图确定。测试系统在 147 只南方肉羊品种上进行了测试。所开发的 DGKH 和 PPP1R1C 基因测试系统可通过 PCR-RT 明确确定绵羊的基因型。在 DGKH 基因中发现了所有三种基因型(T/T、C/T 和 C/C)。在所研究的群体中,没有在 PPP1R1C 基因中发现同源基因型(T/T)。测试系统适用于分子遗传实验室的常规 DNA 分析。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development and validation of a test system for determining the polymorphism in the DGKH and PPP1R1C genes associated with body weight of sheep
Relevance. National sheep breeding is lagging behind other branches of animal husbandry considering using of modern DNA technologies. The search for new genes — potential candidates associated with economically significant traits — is relevant for a more complete disclosure of the genetic potential of sheep. An earlier genome-wide association study showed that the DGKH and PPP1R1C genes had a certain effect on body weight of sheep at birth and at the age of 90 days. In this regard, a more detailed study of polymorphism in the DGKH and PPP1R1C genes can deepen understanding of growth and development processes in domestic sheep, so we have chosen these genes as targets for the experiment.Methods. Primers and probes were selected for amplification of a fragment with targeted SNPs in the DGKH and PPP1R1C genes with a length of 68 nucleotide pairs based on the reference DNA sequence on the 10th (NC_056063.1) and the 2nd chromosome (NC_056055.1) sheep represented in NCBI. To determine polymorphism, real-time PCR-based test systems were developed. Genotypes were determined using a multiparametric graph. The test systems were tested on 147 sheep of the southern meat breed.Results. The developed test systems for promising DGKH and PPP1R1C genes allowed to clearly determine sheep genotypes based on using the PCR-RT. All three genotypes (T/T, C/T, and C/C) were identified in the DGKH gene. No homozygous genotype (T/T) was identified in the PPP1R1C gene in the studied population. The test systems are suitable for routine DNA analysis in molecular genetic laboratories.
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