Exploring DNA Polymorphisms of Leptin Gene within Indian Water Buffaloes
Keywords:Leptin; DNA polymorphism; Buffaloes; PCR-RFLP; SSCP; DNA Sequencing
Leptin is a 16-kDa protein secreted predominantly from white adipose tissue and performs important roles in controlling the body weight, feed intake, immune function, production and reproduction. In present study, buffalo leptin gene was analyzed using DNA markers for polymorphism and its association with production traits. The PCR-RFLP analysis of leptin gene using BsaAI, Sau3AI and Kpn2I restriction enzymes revealed monomorphism. The PCR-SSCP followed by DNA sequencing of Murrah buffalo leptin gene spanning exon III revealed two SNPs viz., c.25 T>C and c.316A>G, which were used to genotype the Murrah, Surti and Bhadawari populations. The c.25 T>C allele frequencies was found 0.81 and 0.19 for C and T in Murrah; 0.65 and 0.35 for C and T in Surti and 0.65 and 0.35 for C and T in Bhadawari buffaloes respectively. The c.25T>C genotypes were not in Hardy-Weinberg proportions, however c.316A>G genotypes maintained equilibrium at this locus in the populations studied. The c.316A>G SNP allele frequencies were observed in Murrah 0.31 and 0.69 for A and G; Surti 0.3 and 0.7 for A and G and Bhadawari 0.3 and 0.7 for A and G respectively. The Murrah buffalo leptin allelic variant sequences revealed 99 % similarity with cattle sequence. The statistical analysis using general linear model procedure indicated that Murrah buffalo leptin c.25 T>C and c.316A>G SNP genotypes were not related significantly (P>0.01) with Murrah buffalo milk production traits viz; milk yield, fat percentage and SNF percentage.
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