Diacylglycerol acyltransferases (DGAT) are key enzymes in fat storage, converting diacylglycerol and fatty acyl CoA into triacylglycerol. In cattle (Bos taurus), the DGAT1 and DGAT2 genes are well-known for their significant influence on milk production traits, particularly milk fat yield and percentage. However, the cattle genome contains twelve other DGAT gene family members that remain largely uncharacterized. The research examined the genetic makeup of these DGAT proteins, revealing differences in exon count, isoforms, amino acid composition, molecular weight, isoelectric points, and predicted SNP locations. The genes are distributed across five chromosomes (2, 14, 15, 25, and X), and the expansion of this gene family in cattle is likely the result of gene duplication events, driven by specific motifs that favor such duplication. By identifying these distinctions, the study provides foundational insights into the lesser-known DGAT genes, which could be involved in regulating important traits like milk fat production. This comprehensive in silico analysis of the DGAT gene family offers valuable insights into the genetic and structural diversity of these enzymes in cattle. It also establishes a platform for future research into functional SNPs and their potential role in improving milk fat production traits, contributing to the advancement of cattle genetics and dairy production optimization. To this end, the identification of previously uncharacterized DGAT genes and their possible involvement in milk fat synthesis could pave the way for innovative breeding strategies focused on improving both milk yield and fat content in dairy cattle.