Polymorphism study of TLR4 gene in crossbred cattle of Kerala

Abstract

The present study was conducted to identify single nucleotide variations in exon 3 region of the bovine Toll-Like Receptor gene (TLR4) associated with somatic cell score (SCS) in crossbred cattle of Kerala using polymerase chain reaction-single stranded conformational polymorphisms (PCR-SSCP) technique. Blood samples were collected from 200 lactating crossbred cattle in different farms of Kerala Veterinary and Animal Sciences University (KVASU) and field centres of ICAR- FPT scheme, KVASU. Genomic DNA was isolated and polymorphisms were identified by SSCP analysis. The 231 bp fragment of exon 3 in TLR4 gene was found to polymorphic and two genotypes viz. CC and CD were obtained with frequencies 77 per cent and 23 per cent respectively. The frequencies of C and D allele in crossbred were found to be 0.88 and 0.12. On sequencing, C to T transition had led to one non-synonymous change at 2021th position of ORF. Cattle with CC genotype were associated with significantly lower SCS (p≤0.05) as compared to CD genotype. The association between TLR4 polymorphism and SCS described in this study suggests the possible role of SNP during mastitis infection. Thus, could be employed as a useful marker for genetic selection to promote mastitis resistance in dairy cattle.

Keywords

• tlr4
• pcr-sscp
• somatic cell score
• crossbred cattle