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Research
(Published
online: 04-12-2014)
2. Cluster of differentiation 14 gene
polymorphism and its association with incidence of clinical
mastitis in Karan fries cattle -
A. Sakthivel Selvan,
I. D. Gupta, A. Verma, M. V. Chaudhari and V. Kumar
Veterinary World, 7(12): 1037-1040
doi:
10.14202/vetworld.2014.1037-1040
A.
Sakthivel Selvan:
Molecular Genetics Lab, Dairy Cattle Breeding Division, National
Dairy Research Institute, Karnal - 132 001, Haryana, India;
drasakthivel1987@gmail.com
I. D.
Gupta:
Molecular Genetics Lab, Dairy Cattle Breeding Division, National
Dairy Research Institute, Karnal - 132 001, Haryana, India;
idgupta1959@gmail.com
A.
Verma:
Molecular Genetics Lab, Dairy Cattle Breeding Division, National
Dairy Research Institute, Karnal - 132 001, Haryana, India;
archana.ndri@gmail.com
M. V.
Chaudhari:
Molecular Genetics Lab, Dairy Cattle Breeding Division, National
Dairy Research Institute, Karnal - 132 001, Haryana, India;
mvet99@gmail.com
V.
Kumar: Molecular Genetics Lab, Dairy Cattle Breeding Division,
National Dairy Research Institute, Karnal - 132 001, Haryana,
India;
vetvkt1986@gmail.com
Received:
06-08-2014, Revised: 04-11-2014, Accepted: 07-11-2014, Published
Online: 04-12-2014
Corresponding author:
I. D. Gupta, e-mail: idgupta1959@gmail.com
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 (χ2) 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.
Keywords: cluster of differentiation 14, Hinf1, Karan
Fries, mastitis, restriction fragment length polymorphism, single
nucleotide polymorphism.
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