Analysis of the Nebulin-Related Anchoring Protein Gene (N-RAP) SNP Polymorphism (C/T) in Slovak Warmblood Horse by PCR-RFLP Method
Keywords:
N-RAP gene, PCR-RFLP, show-jumping, Slovak warmblood horseAbstract
The show-jumping is a important breeding goal in many breeds of warmblood horses. By using of genome-wide association studies (GWAS) were discovered a several genes which could be identified as candidate genes for physical performance in show jumping of horses. The one of this genes is the nebulin-related anchoring protein gene (NRAP) localized on ECA1. The aim of this work was genotyping the Slovak warmblood horse for SNP mutation (rs68545483) in NRAP gene and analysis of genotype structure in population of Slovak warmblood horses. The SNP polymorphism C/T of the N-RAP gene was studied in a group of 40 animals. Genomic DNA was isolated from samples of blood by using salting-out method. The SNP C/T was analyzed by PCR-RFLP method with restriction endonuclease BseGI. The allele T was detected by two restriction fragments 93 bp and 40 bp and the allele C with 133 bp fragment. In the analyzed population of Slovak warmblood horses were detected the following frequency of alleles and genotypes for the SNP polymorphisms C/T of the N-RAP gene. Frequencies of allele T and allele C were 0.6125 and 0.3875 and frequencies of genotypes were 0.15 (genotype CC), 0.475 (genotype CT) and 0.375
(genotype TT).
References
Schrȍder, W., Klostermann, A., Stock, F.K., Distl, O., A genome wide association study for quantitative trait loci of show-jumping in Hanoverian warmblood horses, Animal Genetics, 2011. doi: 10.1111/j.1365-2052.2011.02265.x
Barrey, E., Review: Genetics and genomics in equine exercise physiology: an overview of the new applications of molecular biology as positive and negative markers of performance and health, Equine vet. J., 2010, doi: 10.1111/j.2042-3306.2010.00299.x
Bray, M.S., Hagberg, J.M., Pérusse, L., Rankinen, T., Roth, S.M., Wolfarth, B., Bouchard, C., The human gene map for performance and health-related fitness phenotypes: the 2006-2007 update. Med. Sci. Sports Exerc., 2009, 41, 35-73
Luo, G., Zhang, J.Q., Nguyen, T.P., Herrera, A.H., Paterson, B., Horowits, R., Complete cDNA sequence and tissue localization of N-RAP, a novel nebulin-related protein of striated muscle, Cell Motil.Cytoskel., 1997, 38, 75-90
Carroll, S.L., Herrera, A.H., Horowits, R., Targeting and functional role of N-RAP, a nebulin-related LIM protein, during myofibril assembly in cultured chick cardiomyocytes, J. Cell Sci., 2001, 114, 4229-4238.
Carroll, S., Lu, S., Herrera, A. H., Horowits, R., N-RAP scaffolds I-ZI assembly during myofibrillogenesis in cultured chick cardiomyocytes, J Cell Sci., 2004, 117, 105–114
Dhume, A., Lu, S., Horowits, R., Targeted disruption of N-RAP gene function by RNA interference: A role for N-RAP in myofibril organization, Cell Motil Cytoskeleton, 2006, 63, 493–511
Carroll, S.L., Horowits, R., Myofibrillogenesis and formation of cell contacts mediate the localization of N-RAP in cultured chick cardiomyocytes, Cell Motil Cytoskeleton, 2000, 47, 63–76
Lu, S., Carroll, S.L., Herrera A.H., Ozanne, B., Horowits, R., New NRAP-binding partners a-actinin, filamin and Krp1 detected by yeast two-hybrid screening: Implications for myofibril assembly, J Cell Sci., 2003, 116, 2169–2178
Lu, S., Borst, D.E., Horowits, R., N-RAP expression during mouse heart development. Dev Dyn, 2005, 233, 201–212.
Frank, M.Y., Naxin, H., Yong, Q.G., Ming-cheng, L., Yaqin, M., Dave, H., Gerard, R.L., Jan, D. and Olin D, A. BatchPrimer3: a high throughput web application for PCR and sequencing primer designi. BMC Bioinformatics, 2008, doi:10.1186/1471-2105-9-253
Hill, E.W., Gu, J., Eivers, S.S., Fonseca, R.G., McGivney, B.A., Govindarajan, P., Orr, N., Katz, L.M., MacHugh, D.A., Sequence polymorphism in MSTN predicts sprinting ability and racing stamina in Thoroughbred horses. PLoS ONE, 2010, 5, e8645.