Instability of plasmid DNA sequences: Macro and micro evolution of the antibiotic resistance plasmid R6-5 |
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Authors: | Kenneth N Timmis Felipe Cabello Isabel Andrés Alfred Nordheim Hans J Burkhardt and Stanley N Cohen |
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Institution: | (1) Department of Medicine, Stanford University School of Medicine, 94305 Stanford, California, USA;(2) Department of Genetics, Stanford University School of Medicine, 94305 Stanford, California, USA;(3) Max-Planck-Institut für Molekulare Genetik, D-1000 Berlin 33;(4) Present address: Department of Microbiology, New York Medical College, 10595 Valhalla, New York, USA;(5) Lenrstuhl für Mikrobiologie, Universität Erlangen-Nürnberg, Erlangen, Germany |
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Abstract: | Summary Detailed examination of the structure of cloned DNA fragments of the R6-5 antibiotic resistance plasmid has revealed a substantial degree of polynucleotide sequence heterogeneity and indicates that sequence rearrangements in plasmids and possibly other replicons occur more frequently than has hitherto been appreciated. The sequence changes in cloned R6-5 fragments were shown in some instances to have occurred prior to cloning, i.e. existed in the original population of R6-5 molecules that was obtained from a single bacterial clone and by several different criteria judged to be homogeneous,and in others to have occurred either during the cloning procedure or during subsequent propagation of hybrid molecules. The molecular changes that are described involved insertion/deletion of the previously characterized IS2 insertion element, formation of a new inverted repeat structure probably by duplication of a preexisting R6-5 DNA sequence, sequence inversion, and loss and gain of restriction endonuclease cleavage sites. |
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