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August 1, 1972Proceedings of the National Academy of Sciences2,731 citations

Nonchromosomal Antibiotic Resistance in Bacteria: Genetic Transformation of Escherichia coli by R-Factor DNA

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SCStanley N. CohenACAnnie ChangLHLeslie Hsu

Key Points

  • Determine whether Escherichia coli can be genetically transformed to express multiple antibiotic resistance via the uptake of purified R-factor plasmid DNA.
  • Treated recipient Escherichia coli cells with calcium chloride (CaCl2) to induce competence for DNA uptake.
  • Introduced purified R-factor DNA in various physical conformations (covalently closed, catenated, nicked open circular, denatured, and sonicated) followed by incubation in drug-free medium before antibiotic challenge.
  • CaCl2-treated E. coli took up purified R-factor DNA, expressing immediate drug resistance in a subset of cells and establishing full phenotypic resistance following incubation in drug-free medium.
  • Transformed bacteria stably acquired a closed circular, transferable DNA species matching the resistance profile, fertility, and sedimentation properties of the parent R factor.
  • Covalently closed, catenated, and open circular forms of R-factor DNA all successfully transformed recipient cells, whereas denaturation and sonication completely abolished transformation ability.

Abstract

Transformation of E. coli cells treated with CaCl(2) to multiple antibiotic resistance by purified R-factor DNA is reported. Drug resistance is expressed in a small fraction of the recipient bacterial population almost immediately after uptake of DNA, but full genetic expression of resistance requires subsequent incubation in drugfree medium before antibiotic challenge. Transformed bacteria acquire a closed circular, transferable DNA species having the resistance, fertility, and sedimentation characteristics of the parent R factor. Covalently-closed, catenated, and open (nicked) circular forms of R-factor DNA are all effective in transformation, but denaturation and sonication abolish the transforming ability of R-factor DNA in this system.

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Cite This Study

Cohen et al. (1972) studied this question.

synapsesocial.com/papers/69d9446eccb0bba5a5684888https://doi.org/10.1073/pnas.69.8.2110
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