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Gene amplification in Bacillus subtilis.

M Young

    Journal of General Microbiology
    |July 1, 1984
    PubMed
    Summary
    This summary is machine-generated.

    Bacillus subtilis strains carrying a chloramphenicol acetyltransferase gene amplify gene copies when grown with chloramphenicol. This gene amplification offers a stable, controllable alternative to traditional cloning methods in B. subtilis.

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    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • Bacillus subtilis can be engineered to carry foreign genes, such as the staphylococcal chloramphenicol acetyltransferase gene from pC194.
    • Gene expression and stability in B. subtilis are often influenced by plasmid copy number and integration strategies.

    Purpose of the Study:

    • To investigate the phenomenon of gene copy number alteration in Bacillus subtilis in response to chloramphenicol.
    • To explore the potential of in situ gene amplification as a novel cloning strategy in B. subtilis.

    Main Methods:

    • Culturing a Bacillus subtilis strain containing the staphylococcal chloramphenicol acetyltransferase gene at varying chloramphenicol concentrations.
    • Employing hybridization techniques to quantify the copy number of the amplified DNA sequences per genome.

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    Main Results:

    • Growth in 20 µg/mL chloramphenicol induced a 15-fold amplification of the chloramphenicol acetyltransferase gene sequences.
    • Absence of chloramphenicol led to a loss of these gene sequences.
    • The mechanism appears similar to 'R factor transitioning'.

    Conclusions:

    • In situ gene amplification in B. subtilis provides a controllable and stable method for maintaining cloned sequences.
    • This approach offers advantages over conventional cloning strategies reliant on autonomous plasmids.
    • Hybridization methods used are valuable for determining plasmid copy numbers.