Two-dimensional analysis of head extracts from Drosophila melanogaster identified the four eye-specific protein spots corresponding to the retinin protein. The retinin protein spots were specifically stained with phosphoprotein-specific dye, suggesting that the retinin protein undergoes post-translational modification by phosphorylation. Northern blot analysis showed that the retinin gene begins to be expressed during the late stage of puparium formation during development. Analysis of the N-terminal sequence and expression of the retinin gene in S2 suggest that retinin is a secretory protein. Transgenic flies with knockdown expression of the retinin gene by RNA interference (RNAi) were established. However, no significant phenotypic changes in eye structure or phototransduction were observed in the transgenic flies. Western blot analysis and immunohistochemical studies of D. melanogaster eyes suggest that retinin is a cornea-specific protein.