Phylogeny of cytokines: molecular cloning and expression analysis of sea bass Dicentrarchus labrax interleukin-1beta.
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.
Scapigliati
G
G
Dipartimento di Scienze Ambientali, Università della Tuscia, Viterbo, Italy. scapigg@unitus.it
Buonocore
F
F
Bird
S
S
Zou
J
J
Pelegrin
P
P
Falasca
C
C
Prugnoli
D
D
Secombes
C J
CJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001492:Bass / Q000235:genetics / Q000276:immunology*; D015152:Blotting, Northern; D002478:Cells, Cultured; D003001:Cloning, Molecular; D016207:Cytokines / Q000145:classification / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry; D005784:Gene Amplification; D007375:Interleukin-1 / Q000737:chemistry / Q000145:classification* / Q000235:genetics*; D007962:Leukocytes / Q000276:immunology* / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment / Q000662:veterinary; D020539:Sequence Analysis, Protein / Q000662:veterinary; D017385:Sequence Homology; D013045:Species Specificity
D016207:Cytokines; D018076:DNA, Complementary; D007375:Interleukin-1