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Despite recent advances in our ability to genetically manipulate  Rickettsia , little has been done to employ genetic tools to study the expression and localization of  Rickettsia  virulence proteins. Using a  mariner -based  Himar1  transposition system, we expressed an epitope-tagged variant of the actin polymerizing protein RickA under the control of its native promoter in  Rickettsia parkeri , allowing the detection of RickA using commercially-available antibodies. Native RickA and epitope-tagged RickA exhibited similar levels of expression and were specifically localized to bacteria. To further facilitate protein expression in  Rickettsia , we also developed a plasmid for  Rickettsia  insertion and expression (pRIE), containing a variant  Himar1  transposon with enhanced flexibility for gene insertion, and used it to generate  R. parkeri  strains expressing diverse fluorescent proteins. Expression of epitope-tagged proteins in  Rickettsia  will expand our ability to assess the regulation and function of important virulence factors.

Expression of an Epitope-Tagged Virulence Protein in Rickettsia parkeri Using Transposon Insertion