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A method for isolation of protoplasts from dermatophytes.

T Srikantha, G R Rao

    Journal of General Microbiology
    |June 1, 1984
    PubMed
    Summary

    Researchers developed a new method to isolate protoplasts from dermatophytes using Novozym 234 and MgSO4 flotation. This technique yields cleaner protoplasts for higher DNA recovery compared to traditional mycelial methods.

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    Area of Science:

    • Mycology
    • Molecular Biology
    • Biotechnology

    Background:

    • Dermatophytes are fungi that cause skin infections.
    • Efficient isolation of fungal protoplasts is crucial for genetic and molecular studies.
    • Current methods for protoplast isolation can be inefficient or yield contaminated samples.

    Purpose of the Study:

    • To develop and optimize a method for isolating protoplasts from dermatophytes.
    • To evaluate the purity of isolated protoplasts.
    • To compare DNA recovery from protoplasts versus mycelia.

    Main Methods:

    • Protoplast isolation from dermatophytes utilizing Novozym 234.
    • Separation of protoplasts via a flotation technique in magnesium sulfate (MgSO4).
    • Confirmation of cell wall absence using electron microscopy.

    Main Results:

    • Successful isolation of protoplasts from dermatophytes was achieved.
    • Electron microscopy confirmed the complete removal of cell wall material.
    • DNA recovery from isolated protoplasts was significantly higher than from intact mycelia.

    Conclusions:

    • The Novozym 234 and MgSO4 flotation method provides an effective means for isolating pure dermatophyte protoplasts.
    • This optimized isolation technique enhances DNA yield, facilitating downstream molecular applications.
    • The method offers a valuable tool for research involving dermatophyte genetics and cell biology.

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