Osteopontin gene (OPN) is located in the quantitative trait loci (QTL) for milk production traits in bovine chromosome 6. In the present study the impact of cytosine to thymine transition in the intron-4 of OPN gene (g.8514C > T) on milk production traits was analysed in the Holstein Friesian crossbred cattle of Kerala. Genomic DNA was isolated and a fragment of 290 bp enclosing the polymorphic site was amplified and genotyped by restriction fragment length polymorphism (RFLP) using endonuclease, BSeN1. The genetic variants were distributed according to Hardy-Weinberg equilibrium. The T allele was found to be the major one (T/0.76, C/0.24) and the genotype frequencies were TT/0.60, CT/0.32, and CC/0.08. The amplicons of genotypes were sequenced by sangers dideoxy termination technique and confirmed the mutation. Association study using the General Linear Model-Analysis of Variance (GLM-ANOVA) considering marker, season of calving, parity and herd as fixed factors and dairy trait as dependent variable revealed that none of the yield traits (305 day milk yield, peak yield, fat yield, protein yield, solids not fat yield, lactose yield, daily milk yield) or composition traits (fat percent, protein percent, solids not fat percent and lactose percent) of milk production analysed were significantly differed between CT and TT genotyped animals. The OPN gene polymorphism (g.8514C > T) can be suggested for marker assisted selection (MAS) for future breeding programmes for dairy cattle only after extensive association studies.
Innate immune mechanism plays a key role in mammary defense, from recognition of pathogens to activation of nonspecific and specific immunity involved in elimination of pathogens. Expression profiles of innate immune response genes namely Toll like receptor 2 (TLR-2), Peptidoglycan recognition protein 1 (PGLYRP-1), Interleukin 8 receptor (IL-8 R), L-Selectin (SELL), and Osteopontin (OPN) in milk somatic cells of subclinical mastitis (SCM) affected crossbred cows were investigated under this study at transcript level using quantitative real time polymerase chain reaction (qRT-PCR). Dairy cows in mid lactation were screened for SCM using California Mastitis Test (CMT), Somatic Cell Count (SCC) and Electrical Conductivity test (EC). Based on results of SCM screening tests, crossbred cows were clustered into two groups with four Staphylococcus aureus infected SCM cows and four apparently healthy cows. The expressions levels of TLR-2, PGLYRP-1, IL-8 R, SELL, and OPN in milk somatic cells of SCM affected cows were significantly higher (p < 0.05) than healthy cows. These genes could be considered as candidate genes for innate immune response against S. aureus SCM infection.
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