Expression in Escherchia coli and purification of sea bass (Dicentrarchus labrax) interleukin 1beta, a possible immunoadjuvant in aquaculture

Mar Biotechnol (NY). 2004 Jan-Feb;6(1):53-9. doi: 10.1007/s10126-003-0011-y. Epub 2003 Nov 14.

Abstract

Interleukin 1beta (IL-1beta) is a pleiotropic cytokine that plays a pivotal role in regulating immune responses. Our group has recently cloned IL-1beta from sea bass (Dicentrarchus labrax), one of the main Mediterranean aquacultured fish species. The cDNA is 1292 bp and codes for a deduced peptide of 29.4 kDa with a pI of 5.1. As for trout and carp IL-1beta precursor sequence, no candidate cut site for ICE (IL-1beta converting enzyme) enzyme was apparent in the alignments of sea bass IL-1beta with other mammalian IL-1betas. Nevertheless, a possible mature peptide could start at Ala86, giving a protein of 176 amino acids. The nucleotide sequence coding for this polypeptide was cloned into a pQE-30 expression vector. The plasmid was then transformed in Escherichia coli, and the recombinant protein was purified. Finally, we demonstrated that this purified recombinant IL-1beta was able to induce IL-1beta gene expression in a dose-dependent manner on cells purified from sea bass head kidney and could have immunoadjuvant effects in sea bass vaccination experiments.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aquaculture
  • Bass / genetics*
  • Bass / immunology
  • Blotting, Western
  • DNA Primers
  • DNA, Complementary / genetics
  • Electrophoresis, Agar Gel
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli / metabolism
  • Gene Expression Regulation*
  • Immunoglobulins / blood
  • Interleukin-1 / genetics*
  • Interleukin-1 / isolation & purification*
  • Interleukin-1 / metabolism
  • Kidney / metabolism
  • Plasmids / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Vibrio / immunology

Substances

  • DNA Primers
  • DNA, Complementary
  • Immunoglobulins
  • Interleukin-1