Establishment of a replicating plasmid in rickettsia prowazekii

David O. Wood, Andria Hines, Aimee M. Tucker, Andrew Woodard, Lonnie O. Driskell, Nicole Y. Burkhardt, Timothy J Kurtti, Gerald D Baldridge, Ulrike G Munderloh

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Rickettsia prowazekii, the causative agent of epidemic typhus, grows only within the cytosol of eukaryotic host cells. This obligate intracellular lifestyle has restricted the genetic analysis of this pathogen and critical tools, such as replicating plasmid vectors, have not been developed for this species. Although replicating plasmids have not been reported in R. prowazekii, the existence of well-characterized plasmids in several less pathogenic rickettsial species provides an opportunity to expand the genetic systems available for the study of this human pathogen. Competent R. prowazekii were transformed with pRAM18dRGA, a 10.3 kb vector derived from pRAM18 of R. amblyommii. A plasmid-containing population of R. prowazekii was obtained following growth under antibiotic selection, and the rickettsial plasmid was maintained extrachromosomally throughout multiple passages. The transformant population exhibited a generation time comparable to that of the wild type strain with a copy number of approximately 1 plasmid per rickettsia. These results demonstrate for the first time that a plasmid can be maintained in R. prowazekii, providing an important genetic tool for the study of this obligate intracellular pathogen.

Original languageEnglish (US)
Article numbere34715
JournalPloS one
Volume7
Issue number4
DOIs
StatePublished - Apr 17 2012

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Rickettsia prowazekii
plasmids
Plasmids
Pathogens
pathogens
typhus
plasmid vectors
Rickettsia
Epidemic Louse-Borne Typhus
cytosol
Eukaryotic Cells
lifestyle
genetic techniques and protocols
Cytosol
Population
antibiotics
Life Style
Anti-Bacterial Agents

Cite this

Wood, D. O., Hines, A., Tucker, A. M., Woodard, A., Driskell, L. O., Burkhardt, N. Y., ... Munderloh, U. G. (2012). Establishment of a replicating plasmid in rickettsia prowazekii. PloS one, 7(4), [e34715]. https://doi.org/10.1371/journal.pone.0034715

Establishment of a replicating plasmid in rickettsia prowazekii. / Wood, David O.; Hines, Andria; Tucker, Aimee M.; Woodard, Andrew; Driskell, Lonnie O.; Burkhardt, Nicole Y.; Kurtti, Timothy J; Baldridge, Gerald D; Munderloh, Ulrike G.

In: PloS one, Vol. 7, No. 4, e34715, 17.04.2012.

Research output: Contribution to journalArticle

Wood, DO, Hines, A, Tucker, AM, Woodard, A, Driskell, LO, Burkhardt, NY, Kurtti, TJ, Baldridge, GD & Munderloh, UG 2012, 'Establishment of a replicating plasmid in rickettsia prowazekii', PloS one, vol. 7, no. 4, e34715. https://doi.org/10.1371/journal.pone.0034715
Wood DO, Hines A, Tucker AM, Woodard A, Driskell LO, Burkhardt NY et al. Establishment of a replicating plasmid in rickettsia prowazekii. PloS one. 2012 Apr 17;7(4). e34715. https://doi.org/10.1371/journal.pone.0034715
Wood, David O. ; Hines, Andria ; Tucker, Aimee M. ; Woodard, Andrew ; Driskell, Lonnie O. ; Burkhardt, Nicole Y. ; Kurtti, Timothy J ; Baldridge, Gerald D ; Munderloh, Ulrike G. / Establishment of a replicating plasmid in rickettsia prowazekii. In: PloS one. 2012 ; Vol. 7, No. 4.
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