Genetic transformation of the Lyme disease agent Borrelia burgdorferi with coumarin-resistant gyrB.

Abstract

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.

• Authors: D S Samuels, K E Mach, C F Garon
• Journal: Journal of bacteriology (J Bacteriol) Vol. 176 Issue 19 Pg. 6045-9 (Oct 1994) ISSN: 0021-9193 [Print] United States
• PMID: 7928965 (Publication Type: Journal Article)

Chemical References

• Aminocoumarins
• Anti-Bacterial Agents
• Coumarins
• DNA, Bacterial
• Genetic Markers
• Topoisomerase II Inhibitors
• DNA Gyrase
• DNA Topoisomerases, Type II
• coumermycin

Topics

• Aminocoumarins
• Anti-Bacterial Agents (pharmacology)
• Base Sequence
• Borrelia burgdorferi Group (drug effects, enzymology, genetics)
• Coumarins (pharmacology)
• DNA Gyrase
• DNA Topoisomerases, Type II (genetics)
• DNA, Bacterial (genetics)
• Drug Resistance, Microbial (genetics)
• Electroporation
• Genetic Markers (genetics)
• Molecular Sequence Data
• Mutagenesis, Site-Directed
• Restriction Mapping
• Topoisomerase II Inhibitors
• Transformation, Genetic