CONSTRUCTION OF A PROMOTER PROBE VECTOR AUTONOMOUSLY MAINTAINED IN ASPERGILLUS AND CHARACTERIZATION OF PROMOTER REGIONS DERIVED FROM ASPERGILLUS-NIGER AND A-ORYZAE GENOMES

We used a plasmid carrying a sequence for autonomous maintenance in As pergillus (AMA1) and the E. cell uidA gene as a reporter gene to searc h the A. oryzae and A. niger genomes for DNA fragments having strong p romoter activity. beta-glucuronidase (GUS)-producing A. oryzae transfo rmants containing the No. 8AN derived from A. niger, or the No. 9AO de rived from A. oryzae, mere constitutive for the expression of the uidA gene when cultivated in the presence of a variety of: carbon and nitr ogen sources. When the GUS-producing transformants were grown in liqui d culture, the No. 8AN showed an increase of approximately 3-fold in G US activity compared to the amyB (alpha-amylase encoding gene) promote r. There was also a corresponding increase in the amount of GUS gene-s pecific mRNA. When these transformants were grown as rice-koji, the No . 8AN showed an increase of approximately 6-fold compared to the amyB promoter, and the amount of GUS protein produced also increased. These strong promoter regions might be applicable to the production of othe r heterologous proteins in Aspergillus species.