Cluster of differentiation 14 gene polymorphism and its association with incidence of clinical mastitis in Karan fries cattle

Abstract

Aim: The present study was undertaken with the objectives to characterize, identify DNA polymorphism in cluster of differentiation 14 (CD14) gene in Karan Fries (KF) cattle and to analyze association between genetic variants with incidence of clinical mastitis in National Dairy Research Institute (NDRI) herd, Karnal. Materials and Methods:Genomic DNA was extracted using blood of randomly selected hundred KF lactating cattle by phenol-chloroform method. After checking its quality and quantity, polymerase chain reaction (PCR) was carried out using reported primers to amplify 832 base pair region covering nucleotide base position number 1012 to 1843 (part of promoter, 5'UTR, exon 1, intron 1 and part of exon 2) of bovine CD14 gene. The PCR amplified target product was purified, sequenced and further ClustalW analysis was done to align edited sequence with reported Bos taurus sequence (EU148610.1). The restriction fragment length polymorphism (RFLP) analysis was performed for each KF cow using HinfI restriction enzyme (RE). Cows were assigned genotypes obtained by PCR-RFLP analysis and association study was done using Chi-square (χ 2 ) test.Results: After PCR amplification, DNA sequencing of amplicon confirmed the 832 bases covering 1012 to 1843 nucleotide base position of bovine CD14 gene. ClustalW multiple sequence alignment program for DNA revealed six nucleotide changes in KF cows at positions T1117D, T1239G, T1291C, G1359C, G1361A, and G1811A. Cows were also screened using PCR-RFLP with HinfI RE, which revealed three genotypes CC, CD and DD that differed significantly regarding mastitis incidence. Within CC genotype, 72.73% of cows were in a mastitis non-affected group whereas, those in CD and DD genotypes 69.44% and 60.38% respectively were mastitis affected. Conclusion:KF cows with allele C of CD14 gene were less susceptibility to mastitis compared with D allele.

Single nucleotide polymorphism at cluster of differentiation 14 (CD14) gene and its association with fertility traits in crossbred cattle of Kerala

“…They also suggested that -synonymous substitutions exceeding synonymous substitutions indicate the evolution of this protein through positive selection among domestic animals. Selvan et al (2014) screened Karan Fries (KF) cattle of India and reported six nucleotide changes in KF cows at positions T1117D, T1239G, T1291C, G1359C, G1361A and G1811A in a 832bp region (part of promoter, 5'UTR, exon 1, intron 1 and part of exon 2) of bovine CD14. However, Kumar et al (2014) also reported these SNPs except T1117D in Sahiwal (Bos indicus) cows.…”

Molecular characterization and combined genotype association study of bovine cluster of differentiation 14 gene with clinical mastitis in crossbred dairy cattle

“…[ 29 ] where cows with AB and BB genotypes and by Selvan et al . [ 30 ] where cows with CD and DD genotypes were mastitis affected. Further, on comparison within herd (94 animals), combined genotype ABDD was observed having the highest mastitis incidence of 15.96%.…”

Molecular markers for resistance against infectious diseases of economic importance

“…PCR-RFLP with Hinf I enzyme, of CD14 gene (Chromosome 7) by Selvan et al . [ 54 ] in Karan Fries cows revealed three genotypes CC, CD and DD, that differed significantly regarding mastitis incidence. CC genotype was found to be more resistant when compared with CD and DD.…”