Development and validation of a test system for determining the polymorphism in the DGKH and PPP1R1C genes associated with body weight of sheep

Abstract

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.