Open Access
Research
(Published
online: 23-02-2017)
17.
Identification of single nucleotide
polymorphism in protein phosphatase 1 regulatory subunit 11 gene
in Murrah bulls -
Varsha Jain, Brijesh Patel, Farhat Paul Umar, H. M. Ajithakumar,
Suraj K. Gurjar, I. D. Gupta
and Archana Verma
Veterinary World, 10(2): 244-248
doi:
10.14202/vetworld.2017.244-248
Varsha Jain :
Division of
Animal Genetics and Breeding, ICAR-National Dairy Research
Institute, Karnal, Haryana, India.
Brijesh Patel :
Livestock
Production Management Section, ICAR-National Dairy Research
Institute, Karnal, Haryana, India.
Farhat Paul Umar :
Division of Animal
Genetics and Breeding, ICAR-National Dairy Research Institute,
Karnal, Haryana, India.
H. M. Ajithakumar :
Division of Animal
Physiology, ICAR-National Dairy Research Institute, Karnal,
Haryana, India.
Suraj K. Gurjar :
Division of Animal
Physiology, ICAR-National Dairy Research Institute, Karnal,
Haryana, India.
I. D. Gupta :
Division of Animal
Genetics and Breeding, ICAR-National Dairy Research Institute,
Karnal, Haryana, India.
Archana Verma :
Division of Animal
Genetics and Breeding, ICAR-National Dairy Research Institute,
Karnal, Haryana, India.
Received: 24-09-2016, Accepted: 23-01-2017, Published online:
23-02-2017
Corresponding author:
Archana Verma,
e-mail: archana.ndri@gmail.com
Citation:
Jain V, Patel B, Umar FP, Ajithakumar HM, Gurjar SK, Gupta ID,
Verma A (2017) Identification of single nucleotide polymorphism in
protein phosphatase 1 regulatory subunit 11 gene in Murrah bulls,
Veterinary World, 10(2): 244-248.
Abstract
Aim:
This study was
conducted with the objective to identify single nucleotide
polymorphism (SNP) in protein phosphatase 1 regulatory subunit 11
(PPP1R11) gene in Murrah bulls.
Materials and
Methods:
Genomic DNA was
isolated by phenol–chloroform extraction method from the frozen
semen samples of 65 Murrah bulls maintained at Artificial Breeding
Research Centre, ICAR-National Dairy Research Institute, Karnal.
The quality and concentration of DNA was checked by
spectrophotometer reading and agarose gel electrophoresis. The
target region of PPP1R11 gene was amplified using four sets of
primer designed based on Bos taurus reference sequence. The
amplified products were sequenced and aligned using Clustal Omega
for identification of SNPs. Animals were genotyped by polymerase
chain reaction-restriction fragment length polymorphism (PCR-RFLP)
using EcoNI restriction enzyme.
Results:
The sequences
in the NCBI accession number NW_005785016.1 for Bubalus bubalis
were compared and aligned with the edited sequences of Murrah
bulls with Clustal Omega software. A total of 10 SNPs were found,
out of which 1 at 5’UTR, 3 at intron 1, and 6 at intron 2 region.
PCR-RFLP using restriction enzyme EcoNI revealed only AA
genotype indicating monomorphism in PPP1R11 gene of all Murrah
animals included in the study.
Conclusion:
A total
of 10 SNPs were found. PCR-RFLP revealed only AA genotype
indicating monomorphism in PPP1R11 gene of all Murrah animals
included in the study, due to which association analysis with
conception rate was not feasible.
Keywords:
Murrah bulls, protein phosphatase 1 regulatory subunit 11,
polymerase chain reaction, restriction fragment length
polymorphism.
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