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Cloning, subcellular localization and antibacterial functional analysis of NK-lysin in yellow drum (Nibea albiflora).
- Source :
-
Fish & shellfish immunology [Fish Shellfish Immunol] 2023 Oct; Vol. 141, pp. 109061. Date of Electronic Publication: 2023 Sep 06. - Publication Year :
- 2023
-
Abstract
- Vibrio harveyi is the primary pathogenic bacteria affecting Nibea albiflora aquaculture. In a previous phase, our laboratory intentionally exposed N. albiflora to V. harveyi and analyzed the outcomes using a combination of genome-wide association study (GWAS) and RNA-seq. The results revealed that the antimicrobial peptide NK-lysin (YdNkl-1) was a candidate gene for resistance to V. harveyi disease in N. albiflora. To investigate the role of the antimicrobial peptide NK-lysin in N. albiflora's antimicrobial immunity, we screened the YdNkl-1 gene from the transcriptome database. The full-length cDNA of YdNkl-1 gene is 508 bp, with an open reading frame (ORF) of 477 bp, encoding 158 amino acids. The deduced amino acid sequence of YdNkl-1 contains a signal peptide (1st-22nd amino acids) and a Saposin B domain (50th-124th amino acids), akin to mammalian NK-lysin. Phylogenetic tree analysis confirmed that the NK-lysin of teleost fish clustered into a single species, and YdNkl-1 was most closely related to Larimichthys crocea. Subcellular localization showed that YdNkl-1 was distributed in cytoplasm and nucleus of yellow drum kidney cells. Furthermore, YdNkl-1 mRNA transcripts were significantly up-regulated in the skin, gill, intestine, head-kidney, liver, and spleen after V. harveyi infection, suggesting a critical role in N. albiflora's defense against V. harveyi infection. Additionally, we purified and observed the YdNkl-1 protein, which exhibited a potent membrane-disrupting effect on V. harveyi, Pseudomonas plecoglossicida, Vibrio parahaemolyticus, Escherichia coli and Bacillus subtilis. These findings underscore the significance of NK-lysin in N. albiflora's resistance to V. harveyi infection and provide new insights into the crucial role of NK-lysin in the innate immunity of teleost fishes.<br />Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.<br /> (Copyright © 2023 Elsevier Ltd. All rights reserved.)
- Subjects :
- Animals
Phylogeny
Genome-Wide Association Study
Base Sequence
Fish Proteins chemistry
Anti-Bacterial Agents
Fishes genetics
Immunity, Innate genetics
Cloning, Molecular
Antimicrobial Peptides
Mammals metabolism
Perciformes genetics
Perciformes metabolism
Vibrio parahaemolyticus genetics
Fish Diseases
Subjects
Details
- Language :
- English
- ISSN :
- 1095-9947
- Volume :
- 141
- Database :
- MEDLINE
- Journal :
- Fish & shellfish immunology
- Publication Type :
- Academic Journal
- Accession number :
- 37683807
- Full Text :
- https://doi.org/10.1016/j.fsi.2023.109061