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R esearch
(Published online:
09-10-2015)
5. Genotyping of major
histocompatibility complex Class II DRB gene in Rohilkhandi goats
by polymerase chain reaction-restriction fragment length
polymorphism and DNA sequencing -
Kush Shrivastava, Pushpendra Kumar, Nihar Ranjan Sahoo, Amod
Kumar, Mohd. Faheem Khan, Amit Kumar, Arvind Prasad, B. H. M.
Patel, A. Nasir, Bharat Bhushan and Deepak Sharma
Veterinary World, 8(10): 1183-1188
doi:
10.14202/vetworld.2015.1183-1188
Kush
Shrivastava:
Division of Animal Genetics, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; kush_vet@yahoo.co.in
Pushpendra Kumar:
Division of Animal Genetics, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; pushpendra64@gmail.com
Nihar
Ranjan Sahoo:
Division of Animal Genetics, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; vet.nihar@gmail.com
Amod
Kumar:
Division of Animal Genetics, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; amodvet@gmail.com
Mohd.
Faheem Khan:
Division of Animal Genetics, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; faheemphd@gmail.com
Amit
Kumar:
Division of Animal Genetics, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; vetamitchandan07@gmail.com
Arvind
Prasad:
Division of Parasitology, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; drarvindivri@rediffmail.com
B. H.
M. Patel:
LPM Section, Indian Veterinary Research Institute, Izatnagar,
Bareilly, Uttar Pradesh, India; mpatellpm@gmail.com
A.
Nasir:
Division of Parasitology, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; drnasirivri@rediffmail.com
Bharat
Bhushan:
Division of Animal Genetics, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; bhushan.drbharat@gmail.com
Deepak
Sharma: Division of Animal Genetics, Indian Veterinary
Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India;
ds7758@yahoo.co.in
Received: 17-06-2015, Revised: 26-08-2015, Accepted: 05-09-2015,
Published online: 09-10-2015
Corresponding author:
Pushpendra Kumar, e-mail: pushpendra64@gmail.com
Citation:
Shrivastava K, Kumar
P, Sahoo NR, Kumar A, Khan MF, Kumar A, Prasad A, Patel BHM, Nasir
A, Bhushan B, Sharma D (2015) Genotyping of major
histocompatibility complex Class II DRB gene in Rohilkhandi goats
by polymerase chain reaction-restriction fragment length
polymorphism and DNA sequencing, Veterinary World 8(10):
1183-1188.
Abstract
Aim:
To study the major histocompatibility complex (MHC) Class II
DRB1 gene polymorphism in Rohilkhandi goat using polymerase chain
reaction-restriction fragment length polymorphism (PCR-RFLP) and
nucleotide sequencing techniques.
Materials and Methods: DNA was isolated from
127 Rohilkhandi goats maintained at sheep and goat farm, Indian
Veterinary Research Institute, Izatnagar, Bareilly. A 284 bp
fragment of exon 2 of DRB1 gene was amplified and digested using
BsaI and TaqI restriction enzymes. Population genetic parameters
were calculated using Popgene v 1.32 and SAS 9.0. The genotypes
were then sequenced using Sanger dideoxy chain termination method
and were compared with related breeds/species using MEGA 6.0 and
Megalign (DNASTAR) software.
Results: TaqI locus showed three and BsaI locus showed two
genotypes. Both the loci were found to be in Hardy–Weinberg
equilibrium (HWE), however, population genetic parameters suggest
that heterozygosity is still maintained in the population at both
loci. Percent diversity and divergence matrix, as well as
phylogenetic analysis revealed that the MHC Class II DRB1 gene of
Rohilkhandi goats was found to be in close cluster with Garole and
Scottish blackface sheep breeds as compared to other goat breeds
included in the sequence comparison.
Conclusion: The PCR-RFLP patterns showed population to be in
HWE and absence of one genotype at one locus (BsaI), both the loci
showed excess of one or the other homozygote genotype, however,
effective number of alleles showed that allelic diversity is
present in the population. Sequence comparison of DRB1 gene of
Rohilkhandi goat with other sheep and goat breed assigned
Rohilkhandi goat in divergence with Jamanupari and Angora goats.
Keywords: DRB1, major histocompatibility complex Class II,
polymerase chain reaction-restriction fragment length
polymorphism, Rohilkhandi goat, sequencing.
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