Recircularization and Autonomous Replication of a Sheared R-Factor DNA Segment in Escherichia coli Transformants

Controlled shearing of R-factor DNA leads to formation of fragments carrying an antibiotic resistance gene present on, but not expressed by, the intact R-factor. Transformation of CaCl2-treated E. coli by such fragments yields an autonomously replicating tetracycline-resistance plasmid (Tc6-5) that...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1973-05, Vol.70 (5), p.1293-1297
Main Authors: Cohen, Stanley N., Annie C. Y. Chang
Format: Article
Language:English
Subjects:
Antibiotic resistance
Bacteria
Biological Sciences: Biochemistry
Bromides
Calcium Chloride - pharmacology
Centrifugation
Centrifugation, Density Gradient
Cesium
Chlorides
Conjugation, Genetic
DNA
DNA Replication
DNA, Bacterial
DNA, Circular - isolation & purification
Drug Resistance, Microbial
Electron microscopes
Escherichia coli - drug effects
Ethidium
Extrachromosomal Inheritance
Genes
Microscopy, Electron
Molecules
Mutation
Nalidixic Acid - pharmacology
Plasmids
Salmonella - drug effects
Tetracycline - pharmacology
Tetracycline resistance
Transformation, Genetic
Ultracentrifugation
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Summary:Controlled shearing of R-factor DNA leads to formation of fragments carrying an antibiotic resistance gene present on, but not expressed by, the intact R-factor. Transformation of CaCl2-treated E. coli by such fragments yields an autonomously replicating tetracycline-resistance plasmid (Tc6-5) that contains only a small fraction of the genome of the parent R-factor, and lacks both its fertility functions and its other drug-resistance determinants. Although the Tc6-5 plasmid is not self-transmissible, it can interact and/or recombine with conjugally-proficient plasmids that promote its transfer to other bacteria.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.70.5.1293