Rice endosperm plays a very important role in seedling germination and determines the qualities of rice grain. Although studies on specific gene categories in endosperm have been carried out, global view of gene expression at a transcription level in rice endosperm is still limited. To gain a better understanding of the global and tissue-specific gene expression profiles in rice endosperm, a cDNA library from rice endosperm of immature seeds was sequenced. A cDNA array was constructed based on the tentative unique transcripts derived from expression sequence tag (EST) assembling results and then hybridized with cDNAs from five different tissues or organs including endosperm, embryo, leaf, stem and root of rice. Significant redundancy was found for genes encoding prolamin, glutelin, allergen, and starch synthesis proteins, accounting for ∼34% of the total ESTs obtained. The cDNA array revealed 87 significantly expressed genes in endosperm compared with the other four organs or tissues. These genes included 13 prolamin family proteins, 17 glutelin family proteins, 12 binding proteins, nine catalytic proteins and four ribosomal proteins, indicating a complicated biological processing in rice endosperm. In addition, Northern verification of 1,4-alpha-glucan branching enzyme detected two isoforms in rice endosperm, the larger one of which only existed in endosperm.