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Research Article

volume 52 issue 10 (october 2018) : 1409-1412,   Doi: 10.18805/ijar.B-3395

Exploration of genetic polymorphism in targeted region of Protein Phosphatase 1 Regulatory subunit 11 (PPP1R11) in Murrah bulls

V
Varsha Jain
B
Brijesh Patel
I
Ishwar Dayal Gupta
A
Archana Verma
1Division of Animal Genetics and Breeding, ICAR-National Dairy Research Institute, Karnal-132 001, Haryana, India.
Cite article:- Jain Varsha, Patel Brijesh, Gupta Dayal Ishwar, Verma Archana (2017). Exploration of genetic polymorphism in targeted region of Protein Phosphatase 1 Regulatory subunit 11 (PPP1R11) in Murrah bulls. Indian Journal of Animal Research. 52(10): 1409-1412. doi: 10.18805/ijar.B-3395.

ABSTRACT

The present study was carried out with the objective to explore genetic polymorphism in Protein Phosphatase 1 Regulatory subunit 11 (PPP1R11) gene for its possible association with conception rate in Murrah bulls. 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-NDRI, Karnal. The quality and concentration of DNA was checked through spectrophotometer reading. The target region of PPP1R11 gene was amplified using 3 sets of primer designed based on Bos taurus reference sequence. The amplified products were sequenced, edited and aligned using Clustal Omega with Bubalus bubalis for identification of SNPs. A total of 7 SNPs were found, out of which 1 at exon 3 and 6 at 3’UTR. SNP in exon 3 at nt.G1932C resulted in non synonymous (Serine to Threonine) type of amino acid substitution at position 109 of PPP1R11 gene in Murrah. PCR-RFLP was carried out using restriction enzyme BsmAI for genotyping the animals. All Murrah animals included in present study were monomorphic with CC genotype for the targeted region of PPP1R11 gene. 

REFERENCES

  1. Anonymous. (2012). 19th Livestock Census All India Report, DAHD and F, Ministry of Animal Husbandry, Dairying and Fisheries, Ministry of Agriculture, Government of India. PP. 34-35.
  2. BAHS (Basic Animal Husbandry Statistics) (2015). Annual Report. Department of Animal Husbandry, Dairying and Fisheries, Ministry of Agriculture, Government of India.
  3. Cochran, S. D., Cole, J. B., Null, D. J. and Hansen, P.J. (2013). Discovery of single nucleotide polymorphisms in candidate genes associated with fertility and production traits in Holstein cattle. BMC Genetics .14: 49-71. doi:10.1186/1471-2156-14-49.
  4. Khatib, H., Monson, R.L., Huang, W., Khatib, R., Schutzkus, V., Khateeb, H. and Parrish, J.J. (2010). Validation of in vitro fertility genes in a Holstein bull population. J. Dairy Sci. 93, 2244-2249.
  5. Khatib, H. (2015) U.S. Patent No. 9,102,985 B2. Washington, DC: U.S. patent and trade mark office.
  6. Li, G., Penagaricano, F., Weigel, K.A., Zhang, Y., Rosa, G. and Khatib, H. 2012. Comparative genomics between fly, mouse, and cattle identifies genes associated with sire conception rate. J. Dairy Sci. 95(10), 6122–6129.
  7. Mir, M.A., Chakravarty, A.K., Gupta, A.K., Naha, B.C., Jamuna, V., Patil, C.S. and Singh, A.P. (2015). Optimizing age of bull at first use in relation to fertility of Murrah breeding bulls. Vet. World 8(4), 518-522. 
  8. Mumtaz, S. (2016). Identification of Genetic Markers in PPP1R11 Gene and its association with Daughter Pregnancy Rate in Karan Fries Bulls. M.V.Sc.Thesis submitted to ICAR-National Dairy Research Institute (Deemed University), Karnal (Haryana).
  9. Sambrook. J. and Russell, D.W. (2001). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. Website visited: http://www.cirb.res.in (accessed on 25.02.2017)
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    Research Article

    volume 52 issue 10 (october 2018) : 1409-1412,   Doi: 10.18805/ijar.B-3395

    Exploration of genetic polymorphism in targeted region of Protein Phosphatase 1 Regulatory subunit 11 (PPP1R11) in Murrah bulls

    V
    Varsha Jain
    B
    Brijesh Patel
    I
    Ishwar Dayal Gupta
    A
    Archana Verma
    1Division of Animal Genetics and Breeding, ICAR-National Dairy Research Institute, Karnal-132 001, Haryana, India.
    Cite article:- Jain Varsha, Patel Brijesh, Gupta Dayal Ishwar, Verma Archana (2017). Exploration of genetic polymorphism in targeted region of Protein Phosphatase 1 Regulatory subunit 11 (PPP1R11) in Murrah bulls. Indian Journal of Animal Research. 52(10): 1409-1412. doi: 10.18805/ijar.B-3395.

    ABSTRACT

    The present study was carried out with the objective to explore genetic polymorphism in Protein Phosphatase 1 Regulatory subunit 11 (PPP1R11) gene for its possible association with conception rate in Murrah bulls. 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-NDRI, Karnal. The quality and concentration of DNA was checked through spectrophotometer reading. The target region of PPP1R11 gene was amplified using 3 sets of primer designed based on Bos taurus reference sequence. The amplified products were sequenced, edited and aligned using Clustal Omega with Bubalus bubalis for identification of SNPs. A total of 7 SNPs were found, out of which 1 at exon 3 and 6 at 3’UTR. SNP in exon 3 at nt.G1932C resulted in non synonymous (Serine to Threonine) type of amino acid substitution at position 109 of PPP1R11 gene in Murrah. PCR-RFLP was carried out using restriction enzyme BsmAI for genotyping the animals. All Murrah animals included in present study were monomorphic with CC genotype for the targeted region of PPP1R11 gene. 

    REFERENCES

    1. Anonymous. (2012). 19th Livestock Census All India Report, DAHD and F, Ministry of Animal Husbandry, Dairying and Fisheries, Ministry of Agriculture, Government of India. PP. 34-35.
    2. BAHS (Basic Animal Husbandry Statistics) (2015). Annual Report. Department of Animal Husbandry, Dairying and Fisheries, Ministry of Agriculture, Government of India.
    3. Cochran, S. D., Cole, J. B., Null, D. J. and Hansen, P.J. (2013). Discovery of single nucleotide polymorphisms in candidate genes associated with fertility and production traits in Holstein cattle. BMC Genetics .14: 49-71. doi:10.1186/1471-2156-14-49.
    4. Khatib, H., Monson, R.L., Huang, W., Khatib, R., Schutzkus, V., Khateeb, H. and Parrish, J.J. (2010). Validation of in vitro fertility genes in a Holstein bull population. J. Dairy Sci. 93, 2244-2249.
    5. Khatib, H. (2015) U.S. Patent No. 9,102,985 B2. Washington, DC: U.S. patent and trade mark office.
    6. Li, G., Penagaricano, F., Weigel, K.A., Zhang, Y., Rosa, G. and Khatib, H. 2012. Comparative genomics between fly, mouse, and cattle identifies genes associated with sire conception rate. J. Dairy Sci. 95(10), 6122–6129.
    7. Mir, M.A., Chakravarty, A.K., Gupta, A.K., Naha, B.C., Jamuna, V., Patil, C.S. and Singh, A.P. (2015). Optimizing age of bull at first use in relation to fertility of Murrah breeding bulls. Vet. World 8(4), 518-522. 
    8. Mumtaz, S. (2016). Identification of Genetic Markers in PPP1R11 Gene and its association with Daughter Pregnancy Rate in Karan Fries Bulls. M.V.Sc.Thesis submitted to ICAR-National Dairy Research Institute (Deemed University), Karnal (Haryana).
    9. Sambrook. J. and Russell, D.W. (2001). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. Website visited: http://www.cirb.res.in (accessed on 25.02.2017)
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