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Related Experiment Videos

An upstream activating sequence from the Aspergillus nidulans gpdA gene.

P J Punt1, C Kramer, A Kuyvenhoven

  • 1Department of Molecular Genetics and Gene-Technology, Medical Biological Laboratory TNO, Rijswijk, The Netherlands.

Gene
|October 12, 1992
PubMed
Summary

The gpd box, a promoter element from Aspergillus nidulans gpdA gene, significantly boosted reporter gene expression when added to the amdS gene. This enhancement was orientation-dependent and retained normal gene regulation.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The Aspergillus nidulans amdS gene, encoding acetamidase, is highly regulated.
  • The gpdA gene promoter contains a regulatory element known as the gpd box.

Purpose of the Study:

  • To investigate the effect of introducing the gpd box into the amdS gene's upstream region on gene expression.
  • To determine the regulatory properties and potential mechanisms of the gpd box.

Main Methods:

  • Fusion of the lacZ reporter gene to the amdS gene with and without the gpd box.
  • Introduction of the gpd box at various positions and orientations within the amdS upstream region.
  • Analysis of gene expression levels and retention of known regulatory controls (omega-amino acid and carbon catabolite repression).

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Main Results:

  • Introduction of the gpd box increased reporter gene expression up to 30-fold.
  • The observed increase was dependent on the gpd box's orientation and insertion site.
  • Additional gpdA sequences flanking the gpd box diminished its positive effect.
  • The gpd box did not interfere with the regulation of the amdS promoter by AmdR and CreA proteins.

Conclusions:

  • The gpd box possesses at least two distinct activities, one orientation-dependent and another position-dependent near regulatory sites.
  • These activities are likely not involved in the native regulation of the amdS promoter.
  • The gpd box can act as a potent enhancer element for gene expression in A. nidulans.