Cloning and organisation analysis of a hepcidin-like gene and cDNA from Japan sea bass, Lateolabrax japonicus.
A hepcidin gene was amplified from the liver of Lateolabrax japonicus challenged with a mixed bacterial suspension. Using RT-PCR and RACE, a full length cDNA sequence of the hepcidin like antimicrobial peptide was determined. The complete hepcidin cDNA Hepc2 is 581 bases and contains an ORF of 258 bases with a coding capacity of 86 amino acids. The deduced amino acid sequence, which shares eight cysteines at the identical conserved positions, and gene organisation are conserved between Japan sea bass and other fish species. The predicted molecular weight of the peptide is 9.4 kDa. The 3'-untranslated region is composed of 225 bp with a polyadenylation signal AATAAA sequence appearing at 189 nt and the poly(A) tail at 212 nt downstream of stop codon TGA. The predicted signal peptide cleavage site of its deduced peptide is between codons 24 and 25. Japan sea bass hepcidin-like genomic DNA hepc2 sequence including upstream and downstream regions was composed of two introns and three exons. The cloned 173-bp upstream sequence of Japan sea bass hepcidin-like gene contains putative regulatory elements and several binding motifs for transcription factors. High homologies with hepcidin cDNAs and peptides of white bass (Morone chrysops), human and other fish were shown. Hepc2 of Lateolabrax japonicus is a new member of the hepcidin gene family.
Ren
Hong-Lin
HL
State Key Laboratory of Marine Environmental Science, Xiamen University, Siming Road, Xiamen, Fujian 361005, China.
Wang
Ke-Jian
KJ
Zhou
Hong-Ling
HL
Yang
Ming
M
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis / Q000235:genetics*; D001424:Bacterial Infections / Q000662:veterinary; D001483:Base Sequence / Q000235:genetics; D001492:Bass / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000737:chemistry*; D005393:Fish Diseases / Q000382:microbiology; D064451:Hepcidins; D006801:Humans; D008099:Liver / Q000378:metabolism / Q000382:microbiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment / Q000662:veterinary; D017386:Sequence Homology, Amino Acid
D023181:Antimicrobial Cationic Peptides; D017931:DNA Primers; D018076:DNA, Complementary; C578702:HAMP protein, human; D064451:Hepcidins