ISOLATION OF ADENYLATE-CYCLASE GENE-SPECIFIC SEQUENCES FROM OPHIOSTOMA-NOVO-ULMI, CANDIDA-ALBICANS, AND AGARICUS-BISPORUS BY PCR

Degenerate primers corresponding to consensus sequences in the catalyt ic domains of known fungal adenylate cyclases were used to isolate gen e-specific homologs from the Dutch elm disease pathogen Ophiostoma nov o-ulmi, the dimorphic human pathogen Candida albicans, and the commerc ial mushroom Agaricus bisporus. All three fungi gave the expected PCR product of about 390 bp. Computer searches of the databases revealed t hat the products generated from O. novo-ulmi and C. albicans were high ly similar to the adenylate cyclase gene of Magnaporthe grisea, the ri ce blast fungus (91% and 79%, respectively). The PCR product from the homobasidiomycete A. bisporus, on the other hand, showed 78% similarit y to the uac1 gene of the heterobasidiomycete smut fungus, Ustilago ma ydis. Southern hybridization indicated that all three fungi contain a single adenylate cyclase gene. Our data suggest that PCR will be highl y successful for the isolation of adenylate cyclase sequences from oth er fungi.