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Molecular and General Genetics MGG

, Volume 181, Issue 4, pp 424–433 | Cite as

Plasmid transformation in Bacillus subtilis: Fate of plasmid DNA

  • Willem M. de Vos
  • Gerard Venema
  • Umberto Canosi
  • Thomas A. Trautner
Article

Summary

Only multimeric, and not monomeric forms of B. subtilis plasmids can transform B. subtilis cells (Canosi et al. 1978). This finding prompted us to study the physico-chemical fate of plasmid DNA in transformation. Competent cells of B. subtilis were exposed to either unfractionated preparations or to preparations of multimeric plasmid DNA. Plasmid DNA was re-extracted from such cells and then analyzed by sedimentation and isopycnic centrifugation and also defined by its sensitivity to nuclease S1 degradation. No double-stranded plasmid DNA could be recovered from cells transformed with unfractionated plasmid preparations which contained predominantly monomeric covalently closed circular (CCC) DNA, Re-extracted plasmid DNA was single-stranded, had a molecular weight considerably smaller than monomer length DNA and had been subject to degradation to acid soluble products. However, when transformations were performed with multimeric DNA (constructed by in vitro ligation of linearized pC194 DNA), both double-stranded and partially double-stranded DNA could be recovered in addition to single-stranded DNA.

We assume that plasmid DNA is converted to a single-stranded form in transformation, irrespective of its molecular structure. Double-stranded and partially double-stranded DNAs found in transformation with multimeric DNA would be the products of intramolecular annealing.

Keywords

Bacillus Bacillus Subtilis Soluble Product Plasmid Preparation Subtilis Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1981

Authors and Affiliations

  • Willem M. de Vos
    • 1
  • Gerard Venema
    • 1
  • Umberto Canosi
    • 2
  • Thomas A. Trautner
    • 2
  1. 1.Department of Genetics, Biological CentreState University of GroningenHarenThe Netherlands
  2. 2.Abt. TrautnerMax Planck Institut für molekulare GenetikBerlin 33

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