Study of Leptin Gene Polymorphism In Surti and Jaffarabadi Buffaloes by PCR-RFLP

The present research work was undertaken with the objective to study leptin gene polymorphism using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) in Surti and Jaffarabadi buffaloes. Fifty Surti buffaloes maintained at Livestock Research Station under Navsari Agricultural University, Navsari, Gujarat and 50 Jaffarabadi buffaloes from field areas of Junagadh district of Gujarat. Genomic DNA was isolated from 5ml whole blood sample by phenol chloroform method. From the purified genomic DNA, a 522 bp region spanning over a part of intron 2 and exon 3 and a 94 bp region of exon 2 of the leptin gene was amplified using two different set of primers. The 522 and 94 bp PCR products were digested using BsaAI and Kpn2I restriction enzymes, respectively. For the 522 bp PCR leptin fragment, three BsaAI digestion patterns were found in both the breed groups indicating three genotypes; the frequency of AA, AB and BB genotypes were observed as 0.42, 0.52 and 0.06 in Surti and 0.42, 0.44 and 0.14 in Jaffarabadi buffaloes, respectively. In case of 94 bp PCR products, the Kpn2I digestion revealed two alleles (A and B) and two genotypes; The frequency of BB and AB genotypes were 0.96 and 0.03; 0.94 and 0.06 in Surti and Jaffarabadi buffaloes, respectively. We found that the 522 bp leptin gene fragment was fairly polymorphic, while the 94 bp leptin gene was less polymorphic in both buffalo populations under study.