Abstract: Two single spore isolates named PYd21 and PYd15 respectively and their hybrid heterokaryon named H15-21 were studied as materials in this paper. Using sequencing data of genome, transcriptome and digital gene expression profiles of Volvariella volvacea, Phosphoglycerate mutas(PGAM) were cloned and its structure and expression were studied. Alignment of PGAM genes from PYd21 and PYd15 revealed that both sequences to be identical. Structural analysis of PGAM gene revealed that it consisted of 2118bp with an open reading frame(ORF) of 1611bp,encoding a polypeptide of 536 amino acids. PGAM gene contained 10 exons and 9 introns. One type of alternative splicing and 6 alternative splicing sites were identified during RNA processing. TPM(Transcript Per Million Clean Tags) values for PYd21,PYd15 and H15-21 were 35.29,127.91 and 302.89 respectively, which had significant positive correlation to the growth rate of mycelium. The expression levels of PGAM gene in PYd21,PYd15 and H15-21 indicated that two differently type of nucleus in the cell of H1521 interacted each other which affected expression level of PGAM gene in H15-21. The expression levels of PGAM gene were confirmed by real-time quantitative PCR.