A cDNA encoding a novel immune-type receptor (NITR) of Japanese flounder, Paralichthys olivaceus, was isolated from a kidney cDNA library. The cDNA encoded 357 amino acid residues. The amino acid sequence identities between Japanese flounder NITR1 (poNITR1) and previously reported fish NITRs were approximately 30%-40%. The poNITR1 consisted of two extracellular immunoglobulin (Ig) domains (V and V/C2), a transmembrane domain, an immunoreceptor tyrosine-based inhibitor motif (ITIM) and an ITIM-like motif (itim) in the cytoplasmic region. Five potential N-link glycosylation sites (N-X-S/T) are present in the extracellular Ig domains. Seven cysteine (Cys) residues, which are conserved in previously reported NITRs were observed in the extracellular domain of poNITR1. The poNITR1 gene is composed of five exons and four introns spanning approximately 3.4kb. The poNITR1 transcripts were mainly detected in gill, head kidney, trunk kidney, intestine, while it was weakly detected in heart, liver, muscle, peripheral blood leucocytes, skin, spleen and stomach. However, poNITR1 gene expression was not detected in muscle or ovary. NITR gene expression was not induced by LPS or poly I:C. In situ hybridization revealed that Japanese flounder NITR is expressed in both TCR-alpha- and IgM-presenting cells.