Phylogeny of cytokines: molecular cloning and expression analysis of sea bass Dicentrarchus labrax interleukin-1 beta

Giuseppe Scapigliati, F Buonocore, Steven Bird, Jun Zou, P Pelegrin, C Falasca, D Prugnoli, Christopher John Secombes

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Abstract

In this paper the cloning of interleukin-1 beta (IL-1 beta) from the fish Dicentrarchus labrax (sea bass) is described. Using degenerate primers designed from known IL-1 beta sequences, a cDNA fragment was amplified by PCR and elongated by 3' and 5' RACE to give the full-length coding sequence for sea bass IL-1 beta. The cDNA is 1292 bp, lacks a putative ICE cut site, and codes for a deduced peptide of 29.4 kDa with a pI of 5.1. Sequence analysis showed highest amino acid similarity with rainbow trout (62%), Xenopus (46%), and carp (45.5%) IL-1 beta sequences. Expression studies show that sea bass IL-1 beta can be upregulated by bacterial lipopolysaccharide both in vitro and in vivo in leucocytes from blood, head-kidney, spleen, gills and liver, whereas the IL-1 beta transcript was not detectable in thymus and gut-associated lymphoid tissue. Northern blot analysis with head-kidney leucocyte RNA showed a main LPS-upregulated band at 1.3 kb, and two minor bands at 0.9 and 3.0 kb, respectively. Phylogenetic comparisons with IL-1 beta from other vertebrates is presented. (C) 2001 Academic Press.

Original languageEnglish
Pages (from-to)711-726
Number of pages16
JournalFish & Shellfish Immunology
Volume11
Issue number8
DOIs
Publication statusPublished - Nov 2001

Keywords

  • interleukin-1 beta
  • fish
  • sea bass Dicentrarchus labrax
  • cytokines
  • cytokine evolution
  • leucocytes
  • trout oncorhynchus-mykiss
  • sequence
  • cells
  • macrophages
  • alighnment
  • monocytes
  • gene
  • acid

Cite this

Scapigliati, G., Buonocore, F., Bird, S., Zou, J., Pelegrin, P., Falasca, C., Prugnoli, D., & Secombes, C. J. (2001). Phylogeny of cytokines: molecular cloning and expression analysis of sea bass Dicentrarchus labrax interleukin-1 beta. Fish & Shellfish Immunology, 11(8), 711-726. https://doi.org/10.1006/fsim.2001.0347