Peptidoglycan recognition protein-1 gene polymorphism and its association with mastitis in Murrah buffalo

Abstract

A wide variety of environmental or contagious microorganisms implicated in mastitis, impede the economic growth of dairy sector. Identification of polymorphism in candidate gene of host’s immune system and to rule out mastitis resistant allelic form of candidate gene usually remains prime focal point of research.  Bovine peptidoglycan recognition protein-1 (PGLYRP-1), exclusively present in the granules of polymorphonuclear leukocytes has direct microbicidal properties.  The present study was carried out to find the association between PGLYRP-1 polymorphic alleles with mastitis. Milk samples for somatic cell count and blood samples for PCR-RFLP analysis of PGLYRP-1 gene were collected from 20 mastitis negative and 20 mastitis positive Murrah buffaloes. There was significant difference in somatic cell count of mastitis and mastitis free animals. All amplified PCR products of ~862 bp size of partial region of PGLYRP-1 gene were subjected to each restriction enzyme (HincII or TaqαΙ or ApaI). Polymorphism in the partial region of PGLYRP-1 gene had not been established using PCR-RFLP as uniformity in pattern of digested fragments was seen. Target sequence PGLYRP-1 gene of Murrah buffalo was cloned and sequenced. BLAST analysis revealed sequence identity of PGLYRP-1 of Murrah buffalo with Bos taurus (JN085441.1) sequence at NCBI was 96%, 96% with Bos indicus (JN085440.1) and 96% with Bos indicus X Bos taurus (EU746454.1). In phylogenetic tree, the target sequence of PGLYRP-1 gene of Bubalus bubalis are found more closely related to Bos taurus than to Bos indicus.