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. 2000 Jun;182(11):3289-91.
doi: 10.1128/JB.182.11.3289-3291.2000.

Transformation of Rickettsia prowazekii to erythromycin resistance encoded by the Escherichia coli ereB gene

L I Rachek  1 , A Hines, A M Tucker, H H Winkler, D O Wood
Affiliations

Transformation of Rickettsia prowazekii to erythromycin resistance encoded by the Escherichia coli ereB gene

L I Rachek et al. J Bacteriol. 2000 Jun.

Abstract

Rickettsia prowazekii, the etiologic agent of epidemic typhus, is an obligate, intracytoplasmic, parasitic bacterium. Recently, the transformation of this bacterium via electroporation has been reported. However, in these studies identification of transformants was dependent upon either selection of an R. prowazekii rpoB chromosomal mutation imparting rifampin resistance or expression of the green fluorescent protein and flow cytometric analysis. In this paper we describe the expression in R. prowazekii of the Escherichia coli ereB gene. This gene codes for an erythromycin esterase that cleaves erythromycin. To the best of our knowledge, this is the first report of the expression of a nonrickettsial, antibiotic-selectable gene in R. prowazekii. The availability of a positive selection for rickettsial transformants is an important step in the characterization of genetic analysis systems in the rickettsiae.

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Figures

FIG. 1
FIG. 1
Restriction maps of the Madrid E gltA locus (top) and predicted pMW1047 insertion in RPMOB.001 (bottom). H, HindIII; P, PstI; X, XbaI. Numbers below the relevant restriction sites indicate the location of the restriction site on the map. Arrows identify the location and orientation of the gltA and ereB genes.
FIG. 2
FIG. 2
Hybridization of the R. prowazekii gltA gene to a Southern blot of R. prowazekii chromosomal DNA digested with the indicated restriction enzymes and isolated from the Madrid E strain (lanes 1, 3, and 5) or RPMOB.001 (lanes 2, 4, and 6). Molecular size markers (lane 7) are indicated.
See this image and copyright information in PMC

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