Association of IL-8 receptor gene polymorphism with mastitis in Murrah buffalo

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Date
2012
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LUVAS
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Present study was carried out on Interleukin-8 receptor gene of Murrah buffalo with an objective to find out different variants of Interleukin-8 receptor gene & its association with mastitis. Genomic DNA was isolated from randomly selected lactating buffalo maintained at Govt. Livestock Farm, Hisar; Central Institute for Research on Buffaloes, Hisar; Buffalo Research Centre, College of Animal Sciences and Veterinary clinics, LLRUVAS, Hisar by phenol-chloroform isoamylalcohol method. Quality and quantity of DNA was checked and it was found to be of good quality as determined by agarose gel electrophoresis. A published set of primers covering 311 bp nucleotides of IL-8 receptor gene which was reported polymorphic in previous studies was used and PCR products of 311 bp were obtained. PCR-RFLP was done for all the samples from healthy and mastitis animals using Bsp1286I, HpaII, FauI restriction enzymes. Pattern of resolution of digested fragments by all the restriction enzymes was found same i.e xx, zz, and yy respectively in both two groups of animals. All monomorphic bands were found. Target sequence IL-8 receptor gene of Murrah buffalo was sequenced which has been found to span over 313 bp long. Multiple alignments of IL-8 receptor target sequences of Bubalus bubalis between two groups have revealed no variation. Basic Local Alignment Search Tool (BLAST) analysis revealed sequence identity of target region (IL-8 receptor gene) of Murrah buffalo with Bubalus bubalis (AY864732.1) sequence at NCBI was 99%, 99% with Bos taurus (NM_001105038), 98% with Bos indicus (DQ389109.1) and 90-92% with Sus scrofa (XM_003133655.3). No significant association of Interleukin-8 receptor gene with mastitis was found. However there was significant (p<0.05) difference in somatic cell count of healthy (0.40x105 ± 0.4x104) and mastitis animals (35.2x105 ± 46x104). In phylogenetic tree, the target sequence of Interleukin-8 receptor gene of Bubalus bubalis are found more closely related to Bubalus bubalis, Bos taurus than to Sus scrofa and Homo sapiens.
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