Dataset Information

Mapping and genotypic analysis of the NK-lysin gene in chicken.

ABSTRACT: BACKGROUND: Antimicrobial peptides (AMP) are important elements of the first line of defence against pathogens in animals. NK-lysin is a cationic AMP that plays a critical role in innate immunity. The chicken NK-lysin gene has been cloned and its antimicrobial and anticancer activity has been described but its location in the chicken genome remains unknown. Here, we mapped the NK-lysin gene and examined the distribution of a functionally significant single nucleotide polymorphism (SNP) among different chicken inbred lines and heritage breeds. RESULTS: A 6000 rad radiation hybrid panel (ChickRH6) was used to map the NK-lysin gene to the distal end of chromosome 22. Two additional genes, the adipocyte enhancer-binding protein 1-like gene (AEBP1) and the DNA polymerase delta subunit 2-like (POLD2) gene, are located in the same NW_003779909 contig as NK-lysin, and were thus indirectly mapped to chromosome 22 as well. Previously, we reported a functionally significant SNP at position 271 of the NK-lysin coding sequence in two different chicken breeds. Here, we examined this SNP and found that the A allele appears to be more common than the G allele in these heritage breeds and inbred lines. CONCLUSIONS: The chicken NK-lysin gene mapped to the distal end of chromosome 22. Two additional genes, AEBP1 and POLD2, were indirectly mapped to chromosome 22 also. SNP analyses revealed that the A allele, which encodes a peptide with a higher antimicrobial activity, is more common than the G allele in our tested inbred lines and heritage breeds.

SUBMITTER: Lee MO

PROVIDER: S-EPMC4120735 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Mapping and genotypic analysis of the NK-lysin gene in chicken.

Lee Mi Ok MO Yang Ence E Morisson Mireille M Vignal Alain A Huang Yong-Zhen YZ Cheng Hans H HH Muir William M WM Lamont Susan J SJ Lillehoj Hyun Soon HS Lee Sung Hyen SH Womack James E JE

Genetics, selection, evolution : GSE 20140707

<h4>Background</h4>Antimicrobial peptides (AMP) are important elements of the first line of defence against pathogens in animals. NK-lysin is a cationic AMP that plays a critical role in innate immunity. The chicken NK-lysin gene has been cloned and its antimicrobial and anticancer activity has been described but its location in the chicken genome remains unknown. Here, we mapped the NK-lysin gene and examined the distribution of a functionally significant single nucleotide polymorphism (SNP) am ...[more]

PMID: 25001618

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Project description:The synergistic effects of orally-delivered chicken NK-lysin peptide 2 (cNK-2) or recombinant chicken IL-7 (rchIL-7) on vaccination with recombinant Eimeria elongation factor-1α (rEF-1α) against Eimeria maxima (E. maxima) infection was investigated in broiler chickens. Chickens were divided into six groups: control (CON, no Eimeria infection), non-immunized control (NC, PBS), Vaccination 1 (VAC 1, rEF-1α plus cNK-2), Vaccination 2 (VAC 2, rchIL-7 plus cNK-2), Vaccination 3 (VAC 3, rEF-1α/rchIL-7 plus cNK-2), and Vaccination 4 (VAC 4, rEF-1α/rchIL-7 plus cNK-2). All groups, except the CON and NC, were orally treated with cNK-2 for 5 days. The first immunization, except for the VAC 4 group, was performed intramuscularly on day 4, and the second immunization was given with the same concentration of components as the primary immunization one week later. The immunization of the VAC 4 group was carried out by an oral inoculation on the same days. On day 19, all chickens except the CON group, were orally challenged with E. maxima (1.0 × 104 oocysts/chicken). The in vivo vaccination results showed that the VAC 1 and VAC 3 groups produced high (p < 0.05) levels of serum antibody titers to rEF-1α, and the VAC 3 showed enhanced (p < 0.05) levels of serum IL-7. Furthermore, the VAC 3 group showed significantly (p < 0.01) greater body weight gains at 6- and 9-days post-E. maxima infection (dpi) with reduced oocyst shedding at 6 dpi. The average jejunal lesion score of the NC group was 2.5 whereas the VAC 1 group showed a significantly (p < 0.05) lower lesion scores at 6 dpi. E. maxima infection significantly (P < 0.05) up-regulated the expression levels of cytokines (IL-6, IL-10 and IFN-γ) in the jejunum at 4 dpi, but those expressions were down-regulated in VAC 1 or VAC 3 groups. Moreover, the gene expression levels of Jam 2 and Occludin, were significantly (P < 0.05) decreased following E. maxima infection in jejunum at 4 dpi (NC), but their expressions were increased in the VAC 3 group. Collectively, these results showed that the efficacy of rEF-1α vaccination was significantly enhanced when rEF-1α vaccine co-immunized with chIL-7 or cNK-2.

Project description:This study aimed to determine the transcription profile of NK-lysin gene in native chickens. Moreover, it was targeted toward determining the primary, three-dimensional, and molecular dynamic structures of NK-lysin and granulysin peptides to understand their mode of action and intracellular transduction pathways using in silico analysis. The results revealed that NK-lysin gene in native chickens and Gallus gallus were closely related to those of other avian species. However, there was a low sequence homology when aligned with the mammalian peptides. The coding region of NK-lysin peptide in native chickens encoded 140 amino acids as found in G. gallus with a homology of 98% that declined to 20%, particularly in mammalian species. The results revealed that the NK-lysin in native chickens was closely related to that in avian species at a range of 71-76%. However, it was different from that of other mammalians in terms of nucleotide and amino acid identities. The mRNA transcripts of NK-lysin had high and moderate expression levels in the testis and pancreas, respectively. Nonetheless, the small intestine, kidney, spleen, and liver had a low expression level. The NK-lysin peptides contained more than 50% of the total AA with a nonpolar feature, whereas polar AA constituted up to 30% of AA. The results also indicated that the hydrophilic regions and positively charged amino acids were predominant on the surface of the investigated peptides. The NK-lysin was folded in 4-5 helical units and 3-4 loop structures in their saposin domain. The third helical peptide was long in both avian and bovine species (104-123 residues). However, the fourth helical peptide was short in humans, pigs, and chimpanzees (101-123, 104-123, and 102-124 residues, respectively), with the helical unit residues of 95-97, 96-99, and 96-98, respectively. The obtained results can be helpful in developing novel approaches that could be used as alternatives or adjuncts to the existing means of control.

Project description:BackgroundThe chicken karyotype is composed of 39 chromosome pairs, of which 9 still remain totally absent from the current genome sequence assembly, despite international efforts towards complete coverage. Some others are only very partially sequenced, amongst which microchromosome 16 (GGA16), particularly under-represented, with only 433 kb assembled for a full estimated size of 9 to 11 Mb. Besides the obvious need of full genome coverage with genetic markers for QTL (Quantitative Trait Loci) mapping and major genes identification studies, there is a major interest in the detailed study of this chromosome because it carries the two genetically independent MHC complexes B and Y. In addition, GGA16 carries the ribosomal RNA (rRNA) genes cluster, also known as the NOR (nucleolus organizer region). The purpose of the present study is to construct and present high resolution integrated maps of GGA16 to refine its organization and improve its coverage with genetic markers.ResultsWe developed 79 STS (Sequence Tagged Site) markers to build a physical RH (radiation hybrid) map and 34 genetic markers to extend the genetic map of GGA16. We screened a BAC (Bacterial Artificial Chromosome) library with markers for the MHC-B, MHC-Y and rRNA complexes. Selected clones were used to perform high resolution FISH (Fluorescent In Situ Hybridization) mapping on giant meiotic lampbrush chromosomes, allowing meiotic mapping in addition to the confirmation of the order of the three clusters along the chromosome. A region with high recombination rates and containing PO41 repeated elements separates the two MHC complexes.ConclusionsThe three complementary mapping strategies used refine greatly our knowledge of chicken microchromosome 16 organisation. The characterisation of the recombination hotspots separating the two MHC complexes demonstrates the presence of PO41 repetitive sequences both in tandem and inverted orientation. However, this region still needs to be studied in more detail.

Dataset Information

Mapping and genotypic analysis of the NK-lysin gene in chicken.

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Mapping and genotypic analysis of the NK-lysin gene in chicken.

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