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a b s t r a c t Full-length cDNAs of three genes encoding glyceraldehyde-3-phosphate dehydrogenases (GPDs), PoGPD1, PoGPD2-1, and PoGPD2-2, and their corresponding genomic DNA were isolated from the basidiomycete mushroom Pleurotus ostreatus strain H1. The deduced amino acid sequences of PoGPD1, PoGPD2-1, and PoGPD2-2 had a high degree of similarity to known GPDs of other organisms, especially basidiomycetes. Similar to GPDs of other organisms, the three PoGPDs consist of the GPD NAD þ -binding domain, the GPD C-terminal catalytic domain, and conserved amino acid domains responsible for catalysis, cofactor binding, and substrate binding. Genomic sequence analysis revealed that PoGPD1, PoGPD2-1, and PoGPD2-2 contained seven, eight, and seven introns, respectively, with highly conserved positions. Comparison of the genomic sequences of PoGPD2-1 and PoGPD2-2 in the dikaryon of strain H1 and monokaryons of strain H1 revealed that PoGPD2-1 and PoGPD2-2 are different alleles of PoGPD2. Real-time quantitative reverse-transcription-PCR revealed that the expression patterns of PoGPD1 and PoGPD2 differed during fruit-body development. Specifically, transcript levels of PoGPD1 were high in the primordial stage, while transcript levels of PoGPD2 were high in the mycelial stage. Furthermore, levels of both transcripts were the highest in the base of mature fruit bodies.