Cloning, SNP Analysis Of The DGAT1 Gene In Water Buffalo

Diacylglycerol acyltransferase (DGAT1) is considered to be the key enzyme in controlling the synthesis rate of triglycerides. The DGATl gene was recently identified, and DGATl gene became a strong functional candidate gene for milk fat percentage in cattle. The water buffalo DGATl gene was cloned by iterative polymerase chain reaction using PCR primers. A full-length water buffalo DGATl genomic DNA was sequenced (GenBank: AY999090) 12372bp DNA contained 3504bp of 5'region, 568bp of 3'region. Water buffalo DGATl gene has 98, 91, 84 and 82% nucleotide identity in the coding region with bovine, porcine, human and mouse cDNAs, respectively. Water buffalo DGATl gene codes for a protein of 489 amino acides with 98, 91, 85 and 82% identity to bovine, porcine, human and mouse proteins, respectively. In order to detect the polymorphism of water buffalo DGATl gene, we have used single-strand conformation polymorphism (SSCP) analysis. Seven single nucleotide polymorphisms (SNP) were found in a sample of 120 water buffaloes. One of these variants within 17^th exon result in Ala (GCG) →Val (GTG) , six occur within introns else. 900bp of yak DGATl gene was cloned, compared with water buffalo DGATl gene, the sequence has 94% nucleotide identity.