Characterization of 3’ UTR of Nramp1 gene in Murrah buffalo and its association with brucellosis
Abstract
The present investigation was undertaken to study the genetic polymorphism in 3’UTR of Nramp1 gene and exon V-VII of Nramp1 by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique along with nucleotide sequencing and its association with brucellosis in 50 Murrah buffaloes (including 20 Brucella positive animals). PCR were standardized to amplify 3’UTR of Nramp1 gene and exon V-VII Nramp1 using specific primers. When 178 bp PCR products of 3’ UTR of Nramp1 gene were digested with HaeIII, restriction endonuclease, it detected monomorphic genotypic pattern viz. yy, when 959 bp PCR products of Nramp1 gene (exon V-VII) were digested with HaeIII, AluI, RsaI, StyI and TaqI restriction endonucleases, it detected monomorphic genotypic pattern viz. xx, aa, bb, cc, and dd respectively in all the animals under study.In the present study no polymorphism has been revealed,whish can be attributed either to small sample size or absence of SNPs at restriction sites of the restriction endonucleases used in study. However, these observations need to be confirmed on large sample size and more restriction enzymes. The alignment of nucleotide sequence shows that of 3’URT Nramp1 gene was found 99% similar to Bubalus bubalis, 96% similar to Bubalus arnee and 99% similar to Bos taurus. Exon V-VII was found 95% similar to Bubalus bubalis, 94% to Bos taurus and Bos indicus, 89% to Ovis dalli and 87 % to Ovis areis. In phylogenetic tree, our sequence of Nramp1 gene of Bubalus bubalis are found more closely related to Bos indicus and Bos taurus than Ovis dalli and Ovis areis.