aadAConfers Streptomycin Resistance inBorrelia burgdorferi

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Abstract
To enhance genetic manipulation of the Lyme disease spirocheteBorrelia burgdorferi, we assayed theaadAgene for the ability to confer resistance to the antibiotics spectinomycin and streptomycin. Using the previously described pBSV2 as a backbone, a shuttle vector, termed pKFSS1, which carries theaadAopen reading frame fused to theB.burgdorferi flgBpromoter was constructed. The hybridflgBpromoter-aadAcassette confers resistance to spectinomycin and streptomycin in bothB.burgdorferiandEscherichia coli. pKFSS1 has a replication origin derived from the 9-kb circular plasmid and can be comaintained inB.burgdorferiwith extant shuttle vector pCE320, which has a replication origin derived from a 32-kb circular plasmid, or pBSV2, despite the fact that pKFSS1 and pBSV2 have the same replication origin. Our results demonstrate the availability of a new selectable marker and shuttle vector for genetically dissectingB.burgdorferiat the molecular level.