In this study, we identified and cloned the OsEnS38 cDNA sequence in Zhonghua 11 using bioinformatic and RT–PCR analysis. The encoded OsEnS38 contains two conservative cupin_1 domain typical in Bicupin superfamily. Promoter sequence analysis revealed the existence of several confirmed endosperm–specific cis–regulatory elements in the OsEnS38 promoter. It was confirmed by qRT–PCR and in situ hybridization analysis that the expression of OsEnS38 was specifically expression in developing seeds 7–14 days after pollination. Further analysis revealed that OsEnS38 together with its co–expressing genes were liable to participate in post–embryonic development and reproduction. The results suggested that it might play a role in seed development and accumulation of storage.