Deletion analysis of the Taka-amylase A gene promoter using a homologous transformation system in Aspergillus oryzae.
The Taka-amylase A gene (amyB) of Aspergillus oryzae is induced by starch or maltose. The molecular mechanism of the induction was investigated using a fusion of the amyB promoter and the Escherichia coli uidA gene encoding beta-glucuronidase (GUS). To identify the region responsible for high-level expression and regulation within the amyB promoter, a series of deletion promoters was constructed and introduced into the A. oryzae met locus by homologous recombination. Deletion of the region between -377 to -290 (the number indicates the distance in base pairs from the translation initiation point (+1) to the deletion end point) significantly reduced of the GUS activity, but slight reduction of the GUS activity was observed in deletions up to -377. Northern blot analysis showed that reduction of the GUS activity depended upon the expression level of the GUS gene. The region between -377 to -290 is suggested to include the sequence required directly for high-level expression and regulation of the amyB gene.
Tsuchiya
K
K
Research Institute of Brewing Resources Co., Ltd., Tokyo, Japan.
Tada
S
S
Gomi
K
K
Kitamoto
K
K
Kumagai
C
C
Tamura
G
G
D001236:Aspergillus oryzae / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D001709:Biotechnology; D004271:DNA, Fungal / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D017353:Gene Deletion; D005798:Genes, Bacterial; D005800:Genes, Fungal; D005966:Glucuronidase / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014170:Transformation, Genetic; D000516:alpha-Amylases / Q000235:genetics*
D004271:DNA, Fungal; D000516:alpha-Amylases; D005966:Glucuronidase