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 catalytic domains of known fungal adenylate cyclases were used to isolate gene-specific homologs from the Dutch elm disease pathogen Ophiostoma novo-ulmi, the dimorphic human pathogen Candida albicans, and the commercial mushroom Agaricus bisporus. All three fungi gave the expected PCR product of about 390 bp. Computer searches of the databases revealed that the products generated from O. novo-ulmi and C. albicans were highly similar to the adenylate cyclase gene of Magnaporthe grisea, the rice blast fungus (91% and 79%, respectively). The PCR product from the homobasidiomycete A. bisporus, on the other hand, showed 78% similarity to the uac1 gene of the heterobasidiomycete smut fungus, Ustilago maydis. Southern hybridization indicated that all three fungi contain a single adenylate cyclase gene. Our data suggest that PCR will be highly successful for the isolation of adenylate cyclase sequences from other fungi.
Binz
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Department of Botany, University of Toronto at Mississauga, 3359 Mississauga Road North, Mississauga, Ontario L5L 1C6, Canada.
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D000262:Adenylyl Cyclases / Q000235:genetics*; D000364:Agaricus / Q000201:enzymology / Q000235:genetics*; D001203:Ascomycota / Q000201:enzymology / Q000235:genetics*; D002176:Candida albicans / Q000201:enzymology / Q000235:genetics*; D002177:Candidiasis / Q000382:microbiology; D004271:DNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D005800:Genes, Fungal; D006801:Humans; D010935:Plant Diseases / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012984:Software
D004271:DNA, Fungal; D000262:Adenylyl Cyclases