The 5′ flanking region of a glutelin gene was analyzed for interactions with nuclear proteins from immature rice seed. The specific region between positions-272 and -99 was shown to interact with nuclear proteins from immature seeds, but not with those of leaves and roots. Methylation interference experiments revealed that one factor interacted with a specific sequence element between positions -130 and -120 relative to the transcriptional start site. The sequence specificity of this DNA-protein interaction was confirmed by competition experiments using synthetic oligonucleotides. By using a synthetic oligonucleotide as a probe it was also shown that the binding activity was closely correlated with the mRNA levels of this gene during seed maturation. © 1990 Springer-Verlag.