The bacteroides fragilis BtgA mobilization protein binds to the oriT region of pBFTM10

Leonid A. Sitailo, Alexander M. Zagariya, Patrick J. Arnold, Gayatri Vedantam, David W. Hecht

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7 Scopus citations

Abstract

The Bacteroides fragilis conjugal plasmid pBFTM10 contains two genes, btgA and btgB, and a putative oriT region necessary for transfer in Bacteroides fragilis and Escherichia coli. The BtgA protein was predicted to contain a helix-turn-helix motif, indicating possible DNA binding activity. DNA sequence analysis of the region immediately upstream of btgA revealed three sets of inverted repeats, potentially locating the oriT region. A 304- bp DNA fragment comprising this putative oriT region was cloned and confirmed to be the functional pBFTM10 oriT by bacterial conjugation experiments using E. coli and B. fragilis. btgA was cloned and overexpressed in E. coli, and the purified protein was used in electrophoretic mobility shift assays, demonstrating specific binding of BtgA protein to its cognate oriT. DNase I footprint analysis demonstrated that BtgA binds apparently in a single- stranded fashion to the oriT-containing fragment, overlapping inverted repeats I, II, and III and the putative nick site.

Original languageEnglish (US)
Pages (from-to)4922-4928
Number of pages7
JournalJournal of bacteriology
Volume180
Issue number18
StatePublished - Sep 1 1998
Externally publishedYes

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ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Sitailo, L. A., Zagariya, A. M., Arnold, P. J., Vedantam, G., & Hecht, D. W. (1998). The bacteroides fragilis BtgA mobilization protein binds to the oriT region of pBFTM10. Journal of bacteriology, 180(18), 4922-4928.