GENETIC-TRANSFORMATION OF THE LYME-DISEASE AGENT BORRELIA-BURGDORFERIWITH COUMARIN-RESISTANT GYRB

No useful method to genetically manipulate Borrelia burgdorferi, the c ausative agent of Lyme disease, has been developed previously. We have used resistance to the coumarin antibiotic coumermycin A(1), an inhib itor of DNA gyrase, as a genetic marker to monitor the transformation of B. burgdorferi by electroporation. Introduction of site directed mu tations into the gyrB gene demonstrated that transformation was succes sful, provided evidence that homologous recombination occurs on the ch romosome, and established that mutations at Arg-133 of DNA gyrase B co nfer coumermycin A(1) resistance in B. burgdorferi. The coumermycin A( 1)-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.