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Related Experiment Videos

Gene expression from replicating plasmids in Aspergillus nidulans

A Aleksenko1, I Nikolaev, Y Vinetski

  • 1Division of Molecular Genetics, University of Glasgow, UK.

Molecular & General Genetics : MGG
|November 27, 1996
PubMed
Summary
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Extrachromosomal plasmids in Aspergillus enable gene expression and replication. Highly expressed genes on these plasmids can boost the expression of other genes, even without affecting plasmid stability.

Area of Science:

  • Molecular Biology
  • Mycology
  • Genetics

Background:

  • Extrachromosomal replication vectors are crucial tools in fungal genetics.
  • The AMA1 replicator from Aspergillus nidulans facilitates stable plasmid maintenance.
  • Understanding gene expression regulation on episomal vs. chromosomal DNA is essential.

Purpose of the Study:

  • To investigate the replication and expression capabilities of AMA1-based plasmids in Aspergillus.
  • To determine the impact of highly expressed genes on the transcription of other genes within the same plasmid.
  • To compare gene expression and plasmid behavior between episomal and chromosomally integrated systems.

Main Methods:

  • Construction of synthetic plasmids containing argB and inducible beta-galactosidase (bgaS) genes.

Related Experiment Videos

  • Introduction of plasmids into fungal cells for extrachromosomal replication.
  • Monitoring gene expression using Northern hybridization.
  • Analysis of gene expression in both episomal and chromosomally integrated contexts.

    • Main Results:

    • AMA1-based plasmids replicate extrachromosomally in Aspergillus.
    • Transcription of the highly expressed bgaS gene induces transcription of the co-located argB gene, an effect more pronounced on episomal plasmids.
    • Plasmid copy number correlates with argB expression, but bgaS expression levels remain consistent regardless of copy number or integration.
    • High bgaS expression does not compromise plasmid mitotic stability or copy number.

    Conclusions:

    • AMA1 replicator enables functional extrachromosomal plasmids in Aspergillus.
    • Gene expression from multi-copy episomal plasmids can influence transcription of other genes on the same molecule.
    • Episomal expression systems offer a viable alternative to chromosomal integration for gene expression studies in fungi.