Genotyping of calgranulin A gene in Murrah buffaloes by PCR-RFLP

Calgranulin A (S100A8) gene affects the host disease resistance. S100A8 recruits neutrophils and monocytes and activates them to the site of infection to eliminate bacteria more effectively. Present study was undertaken with the objectives to identify polymorphism in Calgranulin A gene and to associate identified genetic variants with the incidence of clinical mastitis in Murrah buffaloes. Genomic DNA was isolated from 100 unrelated lactating Murrah buffaloes by phenol-chloroform extraction method. Three set of primers targeting different regions were used to obtain PCR products at annealing temperatures of 58 degrees C, 63.8 degrees C and 51 degrees C and were of 514 bp, 557 bp and 323 bp respectively. PCR-RFLP analysis was carried out using restriction endonucleases: EcoO109I, PvuII and MboII for contig I, BsmFI and HaeIII for contig II and MboII for contig III. All the three contigs revealed monomorphism. Since, no genetic variants were observed in the animals studied, it was not feasible to analyze for an association with the incidence of mastitis. However, selection favouring a particular allele and also highly conserved nature of the gene is evident.