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Hydrophobins are a family of small, moderately hydrophobic proteins with eight cysteine residues arranged in a conserved pattern. A full-length cDNA, designated Po.hyd, corresponding to a hydrophobin gene of Pleurotus ostreatus was obtained in our previous work. The Po.hyd gene contains a 333 bp open reading frame (ORF), which is interrupted by two typical classI introns. There was no consensus signal for a polyA tail detected in the 3′untranslated region. However, an analogous T- or TG-rich motif was observed that probably influence the formation of the mRNA 3′ end. We assign the putative Po.HYD protein to the classI hydrophobins since its sequence arrangement and hydropathy pattern has a high consensus to other known class I hydrophobins. Northern analysis showed that the Po.hyd gene was abundantly expressed throughout the fruiting process (from primordium to mature fruiting body) but silenced during vegetative growth of the mycelium. Southern blot analysis showed Po.hyd to be a single copy gene in the genome of dikaryotic strain likely to locate at the same locus within the two parental genomes. (© 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)