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Monomer plasmid DNA transforms Streptococcus pneumoniae

C W Saunders, W R Guild

    Molecular & General Genetics : MGG
    |January 1, 1981
    PubMed
    Summary
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    The covalently closed monomer of plasmid pMV158 transforms Streptococcus pneumoniae via two-hit kinetics. Dimer forms showed higher specific activity but were present in lower quantities.

    Area of Science:

    • Molecular Biology
    • Microbial Genetics
    • Plasmid Biology

    Background:

    • Plasmid DNA is crucial for bacterial transformation and genetic exchange.
    • Understanding the physical forms of plasmids and their transforming efficiency is key to microbial genetics.
    • Plasmid pMV158 is a well-characterized molecule used in studies of bacterial transformation.

    Purpose of the Study:

    • To investigate the transforming activity of different forms of plasmid pMV158 in Streptococcus pneumoniae.
    • To determine the kinetics of transformation mediated by monomeric and dimeric forms of covalently closed (CC) plasmid DNA.
    • To correlate the physical properties of plasmid DNA with its biological transforming efficiency.

    Main Methods:

    • Preparative gel electrophoresis to separate plasmid DNA forms.

    Related Experiment Videos

  • Analytical gel electrophoresis and dye-buoyancy gradients to characterize DNA species.
  • Sedimentation velocity analysis to assess DNA molecular weight and conformation.
  • Transformation assays with Streptococcus pneumoniae to measure biological activity.
  • Kinetic analysis (concentration and time dependence) of transformation efficiency.

    • Main Results:

    • The covalently closed (CC) monomer form of plasmid pMV158 demonstrated two-hit kinetics for transforming pneumococcus.
    • A second peak of transforming activity, primarily composed of CC dimer forms, exhibited single-hit kinetics.
    • Monomer CC was identified as the predominant species responsible for two-hit transformation kinetics.
    • Dimeric CC forms possessed higher specific transforming activity compared to monomers, despite being less abundant.

    Conclusions:

    • Plasmid pMV158 transformation in Streptococcus pneumoniae is dependent on the DNA's physical form (monomer vs. dimer).
    • Two-hit kinetics are characteristic of transformation mediated by monomeric CC plasmid DNA.
    • The study elucidates the relationship between plasmid conformation, quantity, and transforming efficiency in bacteria.