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

Fish Shellfish Immunol. 2001 Nov;11(8):711-26. doi: 10.1006/fsim.2001.0347.

Abstract

In this paper the cloning of interleukin-1beta (IL-1beta) from the fish Dicentrarchus labrax (sea bass) is described. Using degenerate primers designed from known IL-1beta 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-1beta. 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-1beta sequences. Expression studies show that sea bass IL-1beta 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-1beta 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-1beta from other vertebrates is presented.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Bass / genetics
  • Bass / immunology*
  • Blotting, Northern
  • Cells, Cultured
  • Cloning, Molecular
  • Cytokines / classification
  • Cytokines / genetics
  • DNA, Complementary / chemistry
  • Gene Amplification
  • Interleukin-1 / chemistry
  • Interleukin-1 / classification*
  • Interleukin-1 / genetics*
  • Leukocytes / immunology*
  • Leukocytes / metabolism
  • Molecular Sequence Data
  • Phylogeny
  • Polymerase Chain Reaction / veterinary
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Sequence Alignment / veterinary
  • Sequence Analysis, Protein / veterinary
  • Sequence Homology
  • Species Specificity

Substances

  • Cytokines
  • DNA, Complementary
  • Interleukin-1