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Identification of a Novel Single Nucleotide Polymorphism in Porcine Beta-Defensin-1 Gene
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 Title & Authors
Identification of a Novel Single Nucleotide Polymorphism in Porcine Beta-Defensin-1 Gene
Pruthviraj, D.R.; Usha, A.P.; Venkatachalapathy, R.T.;
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Porcine beta-defensin-1 (PBD-1) gene plays an important role in the innate immunity of pigs. The peptide encoded by this gene is an antimicrobial peptide that has direct activity against a wide range of microbes. This peptide is involved in the co-creation of an antimicrobial barrier in the oral cavity of pigs. The objective of the present study was to detect polymorphisms, if any, in exon-1 and exon-2 regions of PBD-1 gene in Large White Yorkshire (LWY) and native Ankamali pigs of Kerala, India. Blood samples were collected from 100 pigs and genomic DNA was isolated using phenol chloroform method. The quantity of DNA was assessed in a spectrophotometer and quality by gel electrophoresis. Exon-1 and exon-2 regions of PBD-1 gene were amplified by polymerase chain reaction (PCR) and the products were subjected to single strand conformation polymorphism (SSCP) analysis. Subsequent silver staining of the polyacrylamide gels revealed three unique SSCP banding patterns in each of the two exons. The presence of single nucleotide polymorphisms (SNPs) was confirmed by nucleotide sequencing of the PCR products. A novel SNP was found in the 5`-UTR region of exon-1 and a SNP was detected in the mature peptide coding region of exon-2. In exon-1, the pooled population frequencies of GG, GT, and TT genotypes were 0.67, 0.30, and 0.03, respectively. GG genotype was predominant in both the breeds whereas TT genotype was not detected in LWY breed. Similarly, in exon-2, the pooled population frequencies of AA, AG, and GG genotypes were 0.50, 0.27, and 0.23, respectively. AA genotype was predominant in LWY pigs whereas GG genotype was predominant in native pigs. These results suggest that there exists a considerable genetic variation at PBD-1 locus and further association studies may help in development of a PCR based genotyping test to select pigs with better immunity.
Selection;Disease Resistance;Antimicrobial Peptide;Ankamali;Large White Yorkshire;Polymerase Chain Reaction-Single Strand Conformation Polymorphism [PCR-SSCP];
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Bassam, B. J., G. Caetano-Anolles, and P. M. Gresshoff. 1991. Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal. Biochem. 196:80-83. crossref(new window)

Bagnicka, E., N. Strzalkowska, K. Flisikowski, T. Szreder, A. Jozwik, B. Prusak, J. Krzyzewski, and L. Zwierzchowski. 2007. The polymorphism in the ${\beta}4$-defensin gene and its assotiation with production and somatic cell count in Holstein-Friesian cows. J. Anim. Breed. Genet. 124:150-156. crossref(new window)

Baroncelli, S., E. Ricci, M. Andreotti, G. Guidotti, P. Germano, M. C. Marazzi, S. Vella, L. Palombi, A. De Rossi, and M. Giuliano. 2008. Single-nucleotide polymorphisms in human beta-defensin-1 gene in Mozambican HIV-1-infected women and correlation with virologic parameters. AIDS 22:1515-1517. crossref(new window)

Choi, M. K., M. T. Le, D. T. Nguyen, H. Choi, W. Kim, J. H. Kim, J. Chun, J. Hyeon, K. Seo, and C. Park. 2012. Genome-level identification, gene expression, and comparitive analysis of porcine ${\beta}$-defensin genes. BMC Genetics 13:98.

De, A. K., S. Jeyakumar, A. Kundu, M. S. Kundu, J. Sunder, and M. Ramachandran. 2013. Genetic characterization of Andaman Desi pig, an indigenous pig germplasm of Andaman and Nicobar group of islands, India by microsatellite markers. Vet. Wld. 6:750-753.

Falconer, D. S. and T. F. C. Mackay. 1996. Introduction to Quantitative Genetics, 4th Ed. Longmans Green, Essex, UK.

Ganz, T. 2003. The role of antimicrobial peptides in innate immunity. Integr. Comp. Biol. 43:300-304. crossref(new window)

Hancock, R. E. W. 1997. Peptide antibiotics. Lancet 349:418-422. crossref(new window)

Hasenstein, J. R. and S. J. Lamont. 2007. Chicken gallinacin gene cluster associated with Salmonella response in advanced intercross line. Avian Dis. 51:561-567. crossref(new window)

Jiang, L. H., H. R. Lu, D. X. Huang, J. B. Yi, L. Y. Li, and F. Lin. 2006. Expression of porcine beta-defensin-1 gene in Pichia pastoris. Sheng Wu Gong Cheng Xue Bao. 22:1036-1039.

Jurevic, R. J., M. Bai, R. B. Chadwick, T. C. White, and B. A. Dale. 2003. Single-nucleotide polymorphisms (SNPs) in human beta-defensin-1: high-throughput SNP assays and association with Candida carriage in type I diabetics and nondiabetic controls. J. Clin. Microbiol. 41:90-96. crossref(new window)

Lai, Y. and R. L. Gallo. 2009. AMPed up immunity: how antimicrobial peptides have multiple roles in immune defense. Trends Immunol. 30:131-141. crossref(new window)

Li, C., T. Xu, R. Chen, X. Huang, Y. Zhao, Y. Bao, W. Zhao, and Z. Zheng. 2013. Cloning, expression and characterization of antimicrobial porcine ${\beta}$-defensin-1 in Escherichia coli. Protein Expr. Purif. 88:47-53. crossref(new window)

Sambrook, J. and D. W. Russell. 2001. Molecular Cloning: A Laboratory Manual. 3rd Ed, Cold Spring Harbor Laboratory Press, New York, NY, USA.

Sanger, F., S. Nicklen, and A. R. Coulson. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Nati. Acad. Sci. USA. 74:5463-5467. crossref(new window)

Segat, L., L. Zupin, R. R. Moura, A. V. C. Coelho, B. S. Chagas, A. C. de Freitas, and S. Crovella. 2014. DEFB1 polymorphisms are involved in susceptibility to human papillomavirus infection in Brazilian gynaecological patients. Mem. Inst. Oswaldo. Cruz 109:918-922. crossref(new window)

Shi, J., G. Zhang, H. Wu, C. R. Ross, F. Blecha, and T. Ganz. 1999. Porcine epithelial beta-defensin-1 is expressed in the dorsal tongue at antimicrobial concentrations. Infect. Immun. 67: 3121-3127.

Zhang, G., H. Hiraiwa, H. Yasue, H. Wu, C. R. Ross, D. Troyer, and F. Blecha. 1999. Cloning and characterisation of the gene for a new epithelial beta-defensin. Genomic structure, chromosomal localization, and evidence for its constitutive expression. J. Biol. Chem. 274:24031-24037. crossref(new window)