Research on the Aerial cell-surface Localization of Hydrophobins

疏水蛋白是丝状真菌中发现的一种具有表面活性和自组装能力的小的两性蛋白。为研究疏水蛋白A （HypA）在米曲霉中的定位，构建了HypA -egfp融合基因，并将其导入米曲霉。荧光显微镜下，固体培养的分生孢子表面特异定位荧光。在深层培养中，菌丝几乎没有荧光。采用抗gfp抗体进行western blot分析，在固态培养和深层培养培养基中检测到HypA-eGFP的融合。我们还观察到，在固态培养中，额外的高分子量带被认为是自组装的HypA-eGFP。这表明egfp标记的HypA被正确定位并组装在分生孢子表面。

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.