Sequence specificity of Bacillus subtilis DNA gyrase in vivo

Linearization of pBG0 (a hydrid between Escherichia coli plasmid pBR322 and Staphylococcus aureus plasmid pUB110) was performed by lysis of the oxolinic acid treated Bacillus subtilis protoplasts with sodium dodecyl sulfate. This plasmid DNA linearization was used both for a detailed mapping of DNA...

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Bibliographic Details
Published in:Genetica 1991, Vol.85 (1), p.3-12
Main Authors: BASHKIROV, V. I, ZVINGILA, D. J
Format: Article
Language:English
Subjects:
Bacillus subtilis
Bacillus subtilis - enzymology
Bacillus subtilis - genetics
Base Sequence
Binding Sites - genetics
Biological and medical sciences
Consensus Sequence
DNA Topoisomerases, Type II - genetics
DNA Topoisomerases, Type II - metabolism
DNA, Bacterial - drug effects
DNA, Bacterial - genetics
DNA, Bacterial - metabolism
Fundamental and applied biological sciences. Psychology
Genic rearrangement. Recombination. Transposable element
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Oxolinic Acid - pharmacology
Plasmids - genetics
Recombination, Genetic - genetics
Substrate Specificity
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Summary:Linearization of pBG0 (a hydrid between Escherichia coli plasmid pBR322 and Staphylococcus aureus plasmid pUB110) was performed by lysis of the oxolinic acid treated Bacillus subtilis protoplasts with sodium dodecyl sulfate. This plasmid DNA linearization was used both for a detailed mapping of DNA gyrase cleavage sites of various strength and for the nucleotide sequence determinations at the points of gyrase-mediated scissions by introducing the XhoI linker DNA. A total of 40 plasmids carrying inserted XhoI linker were sequenced by labeling 3' termini of XhoI sites; 38 of them were found to contain a duplication of four base-pairs of the plasmid sequence flanking the linker, which were characteristic of the oxolinic acid-induced DNA cleavage by E. coli DNA gyrase in vitro and in vivo. The relative strength of these sequenced sites was established by comparing their positions to the sites mapped on the appropriate plasmid genome. This allowed us to propose a consensus sequence of B. subtilis DNA gyrase in vivo cleavage site: [sequence: see text] where N is any nucleotide. The bases in parentheses were preferred secondarily. The involvement of DNA gyrase in illegitimate recombination events in Bacillus subtilis is discussed.
ISSN:0016-6707
1573-6857
DOI:10.1007/BF00056101