Some strains of Aspergillus oryzae were shown to have homologs of aflR, a regulatory gene for aflatoxin synthesis in Aspergillus parasiticus. Transcription of an aflR homolog was examined in six strains of A. oryzae having the homologs, using polymerase chain reaction (PCR) coupled with reverse transcription. No PCR product was obtained when the RNA prepared from the A. oryzae strains cultivated under aflatoxin-producing condition was used as template for amplification of the aflR cDNA. By contrast, a PCR product of the expected size was obtained with RNA from A. parasiticus NIAH-26 processed by the same procedure. From genomic DNA of these strains, PCR products of the same size as above were obtained. Possible degradation of the aflR mRNA in the RNA preparation of the A. oryzae strains was negligible, because the calmodulin transcript was detected by PCR from the same RNA samples. Thus, the aflR homologs in the non-aflatoxigenic A. oryzae strains examined are not expressed even under aflatoxin-producing condition.