Expression of an epitope-tagged virulence protein in rickettsia parkeri using transposon insertion

8Citations
Citations of this article
21Readers
Mendeley users who have this article in their library.

Abstract

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.

Cite

CITATION STYLE

APA

Welch, M. D., Reed, S. C. O., Lamason, R. L., & Serio, A. W. (2012). Expression of an epitope-tagged virulence protein in rickettsia parkeri using transposon insertion. PLoS ONE, 7(5). https://doi.org/10.1371/journal.pone.0037310

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free