Directed insertion of a selectable marker into a circular plasmid of Borrelia burgdorferi

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Directed insertion of a selectable marker into a circular plasmid of Borrelia burgdorferi.

Studies of the biology of Borrelia burgdorferi and the pathogenesis of Lyme disease are severely limited by the current lack of genetic tools. As an initial step toward facile genetic manipulation of this pathogenic spirochete, we have investigated gene inactivation by allelic exchange using a mutated borrelial gyrB gene that confers resistance to the antibiotic coumermycin A1 as a selectable m...

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Linear and circular plasmid content in Borrelia burgdorferi clinical isolates.

The genome of Borrelia burgdorferi, the etiologic agent of Lyme disease, is composed of a linear chromosome and more than 20 linear and circular plasmids. Typically, plasmid content analysis has been carried out by pulsed-field gel electrophoresis and confirmed by Southern hybridization. However, multiple plasmids of virtually identical sizes (e.g., lp28 and cp32) complicate the interpretation ...

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Characterization of a circular plasmid from Borrelia burgdorferi, etiologic agent of Lyme disease.

Borrelia burgdorferi, the etiologic agent of Lyme disease, was recently shown to contain plasmid DNA. Two plasmid species have been described in strain CT1, a Wisconsin tick isolate: a 9.2-kilobase entity; and a larger, 70-kilobase entity. Characterization of the 9.2-kilobase entity by using DNase I and restriction endonucleases demonstrated that the plasmid is supercoiled and exists as a stabl...

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Isolation of a circular plasmid region sufficient for autonomous replication and transformation of infectious Borrelia burgdorferi.

Borrelia burgdorferi contains abundant circular and linear plasmids, but the mechanism of replication of these extrachromosomal elements is unknown. A B. burgdorferi 9 kb circular plasmid (cp9) was amplified in its entirety by the polymerase chain reaction and used to construct a shuttle vector that replicates in Escherichia coli and B. burgdorferi. A 3.3 kb region of cp9 containing three open ...

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ژورنال

عنوان ژورنال: Journal of Bacteriology

سال: 1996

ISSN: 0021-9193,1098-5530

DOI: 10.1128/jb.178.20.5946-5953.1996