Cloning and expression analysis of the NFκB inhibitor IκBα of ayu (Plecoglossus altivelis)

Abstract

The NFκB inhibitor (IκBα) is an integral part of NFκB/IκB signaling pathways, which plays roles in a variety of immune responses, such as bacterial infection resistance. By interacting with nuclear transcription factor NFκB, IκBα controls a variety of biological immune gene expressions. In this study, full-length cDNA (1341 bp) of the NFκB inhibitor IκBα (PaIκBα, GenBank Accession No. JN801027) of Plecoglossus altivelis was obtained by RACE and PCR, and included a 5' untranslated region (UTR) (64 bp), a 3' untranslated region (UTR) (341 bp) and an open reading frame (ORF) (936 bp) encoding a polypeptide of 311 amino acids. PaIκBα had high homology with other IκBαs, containing a conserved ankyrin repeat domain, which was required for interacting with NFκB, a PEST sequence in the C-terminus and a signal responsive domain in the N-terminus. The deduced amino acid sequence of PaIκBα shared 95% homology with Osmerus mordax, and 76%, 75%, 70%, and 68% homology with Salmo salar, Oncorhynchus mykiss, Nile tilapia, and Siniperca chuatsi, respectively. Phylogenetic analysis revealed that IκBα of ayu and Osmerus mordax, Salmo salar, Oncorhynchus mykiss, Nile tilapia, and Siniperca chuatsi were in the same phylogenetic tree. RT-PCR analysis showed that PaIκBα mRNA expression was highest in the liver, kidney, intestine, and gills, then followed by the spleen, brain and muscle, and was lowly expressed in the heart. Likewise, after Aeromonas hydrophila infection, the mRNA level of ayu PaIκBα in the liver was also up-regulated.