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New cell fusion method using polymer membrane.

I Karube, E Tamiya, H Matsuoka

    FEBS Letters
    |September 17, 1984
    PubMed
    Summary
    This summary is machine-generated.

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    Researchers used porous polymer membranes for fusing yeast protoplasts. The tetrafluoroethylene (TFE) membrane method achieved a fusion frequency of 1.2 X 10(-6), enabling genetic analysis.

    Area of Science:

    • Biotechnology
    • Microbiology
    • Cell Biology

    Background:

    • Protoplast fusion is a key technique for yeast genetic engineering.
    • Efficient fusion methods are crucial for improving fusion frequency and strain development.

    Purpose of the Study:

    • To evaluate the efficacy of porous polymer membranes for Saccharomyces cerevisiae protoplast fusion.
    • To determine the fusion frequency achieved using nitrocellulose and tetrafluoroethylene (TFE) membranes.

    Main Methods:

    • Protoplasts of Saccharomyces cerevisiae strains AH22 and D13-1A were adsorbed onto nitrocellulose or TFE membranes under slight suction.
    • Protoplast trapping within membrane pores was confirmed via electron microscopy.
    • Membranes with adsorbed protoplasts were incubated on selective media to identify fused colonies.

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  • Fusion was verified using DNA content analysis and genetic markers.

    • Main Results:

    • Several colonies emerged after 5-7 days of incubation at 30°C.
    • The tetrafluoroethylene (TFE) membrane yielded a fusion frequency of 1.2 x 10⁻⁶.
    • Successful fusion was confirmed through genetic and DNA content analyses.

    Conclusions:

    • Porous polymer membranes, particularly TFE, provide an effective platform for yeast protoplast fusion.
    • This method facilitates the generation and selection of fused yeast strains for further research and application.