Deletion analysis of the Taka-amylase A gene promoter using a homologous transformation system in Aspergillus oryzae

Biosci Biotechnol Biochem. 1992 Nov;56(11):1849-53. doi: 10.1271/bbb.56.1849.

Abstract

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.

MeSH terms

  • Aspergillus oryzae / enzymology
  • Aspergillus oryzae / genetics*
  • Base Sequence
  • Biotechnology
  • DNA, Fungal / genetics
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Gene Deletion
  • Genes, Bacterial
  • Genes, Fungal
  • Glucuronidase / genetics
  • Molecular Sequence Data
  • Plasmids
  • Promoter Regions, Genetic
  • Transcription, Genetic
  • Transformation, Genetic
  • alpha-Amylases / genetics*

Substances

  • DNA, Fungal
  • alpha-Amylases
  • Glucuronidase