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Related Experiment Videos

Transformation by integration in Aspergillus nidulans.

J Tilburn, C Scazzocchio, G G Taylor

    Gene
    |December 1, 1983
    PubMed
    Summary

    Researchers achieved DNA-mediated genetic transformation in Aspergillus nidulans using plasmid DNA and polyethylene glycol. This method successfully integrated the acetamidase gene into the fungal genome, enabling genetic modification.

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    Area of Science:

    • Molecular Biology
    • Mycology
    • Genetic Engineering

    Background:

    • DNA-mediated genetic transformation is crucial for studying gene function in fungi.
    • Aspergillus nidulans is a model organism for fungal research.
    • Efficient transformation protocols are needed for genetic manipulation.

    Purpose of the Study:

    • To establish a DNA-mediated genetic transformation protocol for Aspergillus nidulans.
    • To investigate the integration of plasmid DNA into the A. nidulans genome.
    • To optimize transformation frequency using specific plasmid constructs.

    Main Methods:

    • Incubation of Aspergillus nidulans protoplasts with plasmid DNA (pBR322 derivatives) containing the acetamidase gene.
    • Use of polyethylene glycol and CaCl2 to facilitate DNA uptake.

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  • Analysis of transformant strains for gene integration.

    • Main Results:

    • Successful DNA-mediated genetic transformation of A. nidulans was achieved with a frequency of up to 25 transformants per microgram of DNA.
    • Integration of both plasmid (pBR322) and acetamidase gene sequences into the A. nidulans genome was confirmed.
    • No enhancement in transformation frequency was observed with additional ribosomal repeat or mitochondrial origin fragments.

    Conclusions:

    • A robust method for DNA-mediated genetic transformation of Aspergillus nidulans has been developed.
    • Plasmid DNA, including the acetamidase gene, integrates into the host genome.
    • Further optimization may involve exploring different integration sites or regulatory elements.