We constructed a rock bream (Oplegnathus fasciatus) gill cDNA library and a total of 1450 expressed sequence tag (EST) clones were generated. Gene annotation procedures and homology searches of the sequenced ESTs were locally done by BLASTX for amino acid similarity comparisons. Of the 1450 EST clones, 1022 EST clones showed significant homology to previously described genes while 428 ESTs were unidentified, and 259 clones were hypothetical, or unnamed proteins. Encoding 313 different sequences were identified as putative bio-defense genes or genes associated with immune response.