Identification of novel single nucleotide polymorphisms in the DGAT1 gene of buffaloes by PCR-SSCP

Ashwin A Raut,Neeraj Rana,Anil Kumar,Anamika Mishra,Sundeep Jaglan,Vikas Beniwal,Jitender K Singh,Sheo N Kala,Vinod Chhokar,Sachin K Samuchiwal

doi:10.1590/s1415-47572012005000043

Abstract

Diacylglycerol O-acyltransferase 1 (DGAT1) is a microsomal enzyme that catalyzes the final step of triglyceride synthesis. The DGAT1 gene is a strong functional candidate for determining milk fat content in cattle. In this work, we used PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) and DNA sequencing to examine polymorphism in the region spanning exon 7 to exon 9 of the DGAT1 gene in Murrah and Pandharpuri buffaloes. Three alleles (A, B and C) and four novel single-nucleotide polymorphisms were identified in the buffalo DGAT1 gene. The frequencies of the alleles differed between the two buffalo breeds, with allele C being present in Murrah but not in Pandharpuri buffalo. The allele variation detected in this work may influence DGAT1 expression and function. The results described here could be useful in examining the association between the DGAT1 gene and milk traits in buffalo.

Highlights

Diacylglycerol O-acyltransferase 1 (DGAT1) is one of the key enzymes in controlling the rate of triglyceride synthesis in adipocytes
Since the DGAT1 gene maps to the quantitative trait loci (QTL) for milk fat percentage in the centromeric region of chromosome 14 (BTA14), this gene has been studied as a candidate for association with the milk fat content in cattle.Mutation analysis in cattle has revealed 19 polymorphic sites within the DGAT1 gene (Winter et al, 2002)
The aim of this study was to examine the polymorphism in introns 7 and 8 of the DGAT1 gene in Murrah and Pandharpuri buffaloes using polymerase chain reaction-singlestrand conformation polymorphism (PCR-SSCP), a rapid, precise technique that allows the identification of singlenucleotide polymorphisms (SNPs)

Summary

Introduction

Diacylglycerol O-acyltransferase 1 (DGAT1) is one of the key enzymes in controlling the rate of triglyceride synthesis in adipocytes. The aim of this study was to examine the polymorphism in introns 7 and 8 of the DGAT1 gene in Murrah and Pandharpuri buffaloes using polymerase chain reaction-singlestrand conformation polymorphism (PCR-SSCP), a rapid, precise technique that allows the identification of singlenucleotide polymorphisms (SNPs).

Objectives

Results

Conclusion