We constructed a rock bream (Oplegnathus fasciatus) liver cDNA library and a total of 1533 expressed sequence tag (EST) clones were generated. Gene annotation procedures and homology searches of the sequenced ESTs were analyzed using BLASTX. Of the 1533 EST clones, 1165 different ESTs showed significant homology to previously described genes while 368 ESTs were unidentified, hypothetical, or unnamed proteins. Encoding 106 different sequences were identified as putative bio-defense genes or genes associated with immune response.