A cDNA library was constructed from the leaves of the disease-resistant cucumber (Cucumis sativus L.) cultivar ‘D0462’ challenged by Pseudomonas syringae pv. Lachrymans for 48 h. The inserted fragment sizes ranged from 0.45 to 2.1 kb and the average inserted size was 1 kb. Sequencing analysis showed that 2 352 TUTs (Tentative unique transcripts), 282 contigs, and 2 070 singlets were identified in the 2 966 ESTs derived from the cDNA library. The result of the BlastX analysis indicated that there were 1 848 ESTs with known or unknown function, 504 ESTs with no significant similarity matching with any protein or DNA sequence in the databases. In this library, many defense/disease-resistant related genes, such as metallothionein, glutathione S-transferase, ubiquitin, b-1, 3-glucanase, zinc finger protein, and cysteine protease, which might participate in the plant and the pathogens, are inclued.