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Bacterial fusion assayed by a prophage complementation test.

C Sanchez-Rivas, A J Garro

    Journal of Bacteriology
    |March 1, 1979
    PubMed
    Summary
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    This study measured heterozygous fusion products in Bacillus subtilis using prophage complementation. High frequencies were observed, suggesting fusion occurs in two steps and is influenced by growth media.

    Area of Science:

    • Microbiology
    • Bacterial genetics
    • Protoplast fusion

    Background:

    • Previous studies determined cell wall regeneration and recombination frequencies in bacterial protoplast fusion.
    • The frequency of heterozygous fusion products was not previously measured.

    Purpose of the Study:

    • To measure the frequency of heterozygous fusion products in Bacillus subtilis using prophage complementation.
    • To investigate the mechanism and influencing factors of bacterial protoplast fusion.

    Main Methods:

    • Utilized two multiply marked nonsuppressing Bacillus subtilis strains, lysogenic for different Sus mutant phage phi 105.
    • Protoplasts were induced by mitomycin C, fused, and incubated in hypertonic broth.
    • Frequency of heterozygous fused cells determined by infectious centers produced, avoiding cell lysis.

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    Main Results:

    • Observed very high frequencies of heterozygous fusion products, consistent with electron microscopy data.
    • Fusion demonstrated to occur in two steps: one polyethylene glycol dependent, the other energy requiring.
    • Bacterial growth medium inversely affects fusion and cell wall regeneration abilities.

    Conclusions:

    • Prophage complementation is an effective method for determining heterozygous fusion product frequency.
    • Bacterial growth media significantly influence protoplast fusion and regeneration dynamics.
    • An inverse relationship exists between fusion and regeneration abilities, with a direct relationship between heterozygotes and recombinants.