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Genome-level identification, gene expression, and comparative analysis of porcine ß-defensin genes.

ABSTRACT: BACKGROUND: Beta-defensins (?-defensins) are innate immune peptides with evolutionary conservation across a wide range of species and has been suggested to play important roles in innate immune reactions against pathogens. However, the complete ?-defensin repertoire in the pig has not been fully addressed. RESULT: A BLAST analysis was performed against the available pig genomic sequence in the NCBI database to identify ?-defensin-related sequences using previously reported ?-defensin sequences of pigs, humans, and cattle. The porcine ?-defensin gene clusters were mapped to chromosomes 7, 14, 15 and 17. The gene expression analysis of 17 newly annotated porcine ?-defensin genes across 15 tissues using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) showed differences in their tissue distribution, with the kidney and testis having the largest pBD expression repertoire. We also analyzed single nucleotide polymorphisms (SNPs) in the mature peptide region of pBD genes from 35 pigs of 7 breeds. We found 8 cSNPs in 7 pBDs. CONCLUSION: We identified 29 porcine ?-defensin (pBD) gene-like sequences, including 17 unreported pBDs in the porcine genome. Comparative analysis of ?-defensin genes in the pig genome with those in human and cattle genomes showed structural conservation of ?-defensin syntenic regions among these species.

SUBMITTER: Choi MK

PROVIDER: S-EPMC3499285 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Genome-level identification, gene expression, and comparative analysis of porcine ß-defensin genes.

Choi Min-Kyeung MK Le Minh Thong MT Nguyen Dinh Truong DT Choi Hojun H Kim Won W Kim Jin-Hoi JH Chun Jungwan J Hyeon Jiyeon J Seo Kunho K Park Chankyu C

BMC genetics 20121115

<h4>Background</h4>Beta-defensins (β-defensins) are innate immune peptides with evolutionary conservation across a wide range of species and has been suggested to play important roles in innate immune reactions against pathogens. However, the complete β-defensin repertoire in the pig has not been fully addressed.<h4>Result</h4>A BLAST analysis was performed against the available pig genomic sequence in the NCBI database to identify β-defensin-related sequences using previously reported β-defensi ...[more]

PMID: 23150902

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Project description:Backgroundβ-defensins have attracted considerable research interest because of their roles in protecting hosts from various pathogens. This study was conducted to investigate the expression profiles of the porcine β-defensin 114 (PBD114) in different breeds and in response to infections. Moreover, the function of PBD114 protein was partially investigated.MethodsSix Tibetan pigs (TP) and six DLY (Duroc×Landrace×Yorkshire) pigs were slaughtered to explore the expression profiles of PBD114 in different breeds and tissues. For infection models, sixteen DLY pigs were divided into two groups and challenged either with sterile saline or E. coli K88. The recombinant protein PBD114 (rPBD114) was obtained by using a heterologous expression system in E. coli.ResultsPBD114 gene was highly expressed in tissues such as the intestine, liver, spleen, and thymus. Interestingly, the expression level of PBD114 gene was higher in the TP pigs than in the DLY pigs (P < 0.05), and was significantly elevated upon E. coli K88 challenge (P < 0.05). The nucleotide sequences of PBD114 from Tibetan and DLY pigs was identical, and both showed a 210-bp open reading frame encoding a 69-amino acid mature peptide. To explaore the function of PBD114 protein, PBD114 gene was successfully expressed in E. coli Origami B (DE3) and the molecular weight of the rPBD114 was estimated by SDS-PAGE to be 25 kDa. The rPBD114 was purified and mass spectrometry verified the protein as PBD114. Importantly, rPBD114 showed antimicrobial activities against E. coli DH5α and E. coli K88, and the minimal inhibitory concentrations (MICs) were 64 and 128 μg/mL, respectively. Hemolytic and cytotoxicity assays showed that rPBD114 did not affect cell viability under physiological concentrations.ConclusionsPBD114 is an infection response gene that is differentially-expressed between different porcine breeds and tissues. The antimicrobial activity of PBD114 protein, against pathogens such as the E. coli K88, suggested that it may serve as a candidate for the substitution of conventionally used antibiotics.

Dataset Information

Genome-level identification, gene expression, and comparative analysis of porcine ß-defensin genes.

Publications

Genome-level identification, gene expression, and comparative analysis of porcine ß-defensin genes.

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