Characterization of β4-defensin gene and its association with mastitis in Murrah buffaloes
Abstract
The present investigation was undertaken to study the genetic polymorphism in β4-defensin
gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique
along with nucleotide sequencing and its association with mastitis in 60 Murrah buffaloes (including 30
mastitic animals). 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 chloform method. DNA
quality and quantity was checked by agarose gel electrophoresis. PCR was standardized to amplify
partial intron 1 and exon 2 (600 bp) of β4-defensin gene using specific self designed (NCBI pick
primer) primers. When 600 bp PCR products of β4-defensin gene were digested with BfuCI, PstI and
HinfI restriction endonucleases, it detected single genotype viz. xx. Only monomorphism was observed
with all restriction enzymes under study. However there was significant (p<0.05) difference in somatic
cell count of healthy (0.41x105 ± 0.6x104) and diseased animal (26.35x105 ± 40.9x104).
In phylogenetic tree, the β4-defensin gene of Bubalus bubalis are found more closely related
to Bos taurus (96%) than to Capra hircus (89%) and Ovis aries (87%).