Molecular characterization of hepcidin gene in common carp (Cyprinus carpio L.) and its expression pattern responding to bacterial challenge.

Hepcidin is a cysteine-rich cationic antimicrobial peptide (AMP), which plays an important role in host innate immune system and iron regulation. A great many of hepcidin genes have been identified from vertebrates, including various fish species. Here we report the cloning and identification of a hepcidin cDNA from the liver of common carp (Cyprinus carpio L.). The full-length cDNA of the common carp hepcidin was 647 bp, which contained an ORF of 276 bp encoding a prepropeptide of 91 amino acid residues. The predicted preprohepcidin consisted of three domains: a signal peptide of 24 amino acids, a prodomain of 42 amino acids and a mature peptide of 25 amino acids, which containd eight cysteine residues at the identical conserved position. The genomic organization of common carp hepcidin was identified, which contained three exons and two introns, similarly to corresponding genes in mammals and other fish species. Sequence alignment and phylogenetic analysis showed that hepcidins were conserved in different species, and common carp hepcidin was type 1 hepcidin according to the phylogenetic tree, which had the highest identity with mud loach and zebrafish. Real-time PCR assay showed that the hepcidin gene was mostly expressed in liver, and expressed in all the other tested tissues of common carp in different levels. When challenged with pathogenic bacterium, Vibrio anguillarum, the expression level of common carp hepcidin was quickly up-regulated in liver, spleen, head kidney and hindgut, implying that hepcidin may be an important component of the innate immune system of common carp and involved in mucosal immune response against invading pathogens.