Genetic transformation of the Lyme disease agent Borrelia burgdorferi with coumarin-resistant gyrB.
No useful method to genetically manipulate Borrelia burgdorferi, the causative agent of Lyme disease, has been developed previously. We have used resistance to the coumarin antibiotic coumermycin A1, an inhibitor of DNA gyrase, as a genetic marker to monitor the transformation of B. burgdorferi by electroporation. Introduction of site-directed mutations into the gyrB gene demonstrated that transformation was successful, provided evidence that homologous recombination occurs on the chromosome, and established that mutations at Arg-133 of DNA gyrase B confer coumermycin A1 resistance in B. burgdorferi. The coumermycin A1-resistant gyrB marker and genetic transformation can now be applied toward dissecting the physiology and pathogenesis of the Lyme disease agent on a molecular genetic level.
Samuels
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DS
Laboratory of Vectors and Pathogens, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 59840.
Mach
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Garon
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D049933:Aminocoumarins; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001483:Base Sequence; D015748:Borrelia burgdorferi Group / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D003374:Coumarins / Q000494:pharmacology; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics; D018274:Electroporation; D005819:Genetic Markers / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015183:Restriction Mapping; D059005:Topoisomerase II Inhibitors; D014170:Transformation, Genetic
D049933:Aminocoumarins; D000900:Anti-Bacterial Agents; D003374:Coumarins; D004269:DNA, Bacterial; D005819:Genetic Markers; D059005:Topoisomerase II Inhibitors; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II; C004628:coumermycin