Identification of single nucleotide polymorphism in protein phosphatase 1 regulatory subunit 11 gene in Murrah bulls.
OBJECTIVE
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.


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 onBos taurusreference 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 forBubalus bubaliswere 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.


CONCLUSIONS
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.
Division of Animal Genetics and Breeding, ICAR-National Dairy Research Institute, Karnal, Haryana, India.
Varsha
V
Jain
Livestock Production Management Section, ICAR-National Dairy Research Institute, Karnal, Haryana, India.
Brijesh
B
Patel
Division of Animal Genetics and Breeding, ICAR-National Dairy Research Institute, Karnal, Haryana, India.
Farhat Paul
FP
Umar
Division of Animal Physiology, ICAR-National Dairy Research Institute, Karnal, Haryana, India.
H M
HM
Ajithakumar
Division of Animal Physiology, ICAR-National Dairy Research Institute, Karnal, Haryana, India.
Suraj K
SK
Gurjar
Division of Animal Genetics and Breeding, ICAR-National Dairy Research Institute, Karnal, Haryana, India.
I D
ID
Gupta
Division of Animal Genetics and Breeding, ICAR-National Dairy Research Institute, Karnal, Haryana, India.
Archana
A
Verma
Murrah bulls; polymerase chain reaction; protein phosphatase 1 regulatory subunit 11; restriction fragment length polymorphism