Molecular cloning and characterisation of the rock bream, Oplegnathus fasciatus, Fas (CD95/APO-1), and its expression analysis in response to bacterial or viral infection.

Abstract: Fas belongs to the tumour necrosis factor (TNF) receptor superfamily and can transmit a death signal leading to apoptosis. In the present study, we isolated the full-length cDNA for rock bream (Oplegnathus fasciatus) Fas (RbFas). The full-length RbFas cDNA was 1770bp long and contained an open reading frame of 957bp that encoded 319 amino acid residues with a predicted molecular mass of 35.1kDa. The 319 amino-acid predicted RbFas sequence is homologous to other Fas sequences, contains three cysteine-rich domains and a death domain (DD) and two potential N-glycosylation sites. Expression of RbFas mRNA was detected in nine different tissues from healthy rock bream and was the highest in red blood cells. In analyses of mitogen-stimulated RbFas expression in peripheral blood leucocytes, expression of RbFas mRNA was observed between 1 and 36h after stimulation with LPS, and 1 and 3h stimulation with poly I:C. In the case of bacterial injection, the RbFas transcript peaked 6h after injection in both the kidney and the spleen. Otherwise, the RbFas transcript peaked after 1h in spleen and 6h in kidney following injection with RSIV. [Copyright &y& Elsevier]