Research on the Aerial cell-surface Localization of Hydrophobins

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Hydrophobins are small amphipathic proteins found uniquely in filamentous fungi that show surface-activity and self-assembly. To investigate the localization of hydrophobin A (HypA) in Aspergillus oryzae, hypA-egfp fusion gene was constructed and introduced into A.oryzae. By fluorescent microscopy, the fluorescence was specifically localized on surface of conidia in solid-state culture. In submerged cultures, the fluorescence was hardly observed in the hyphae. By western blot analysis using anti-GFP antibody, HypA-eGFP fusion was detected in the solid-state culture and medium of submerged culture. We also observed, an additional high molecular weight band, which was considered to be self-assembled HypA-eGFP, in the solid-state culture. This observation indicates that EGFP-tagged HypA was correctly localized and assembled on the surface of conidia.