Fluorescence-Reported Allelic Exchange Mutagenesis-Mediated Gene Deletion Indicates a Requirement for Chlamydia trachomatis Tarp during <i>In Vivo</i> Infectivity and Reveals a Specific Role for the C Terminus during Cellular Invasion
Susmita Ghosh, Elizabeth A. Ruelke, Joshua Ferrell, Maria D. Bodero, Kenneth A. Fields, Travis J. Jewett
Abstract
The translocated actin recruiting phosphoprotein (Tarp) is a multidomain type III secreted effector used by Chlamydia trachomatis . In aggregate, existing data suggest a role of this effector in initiating new infections. As new genetic tools began to emerge to study chlamydial genes in vivo , we speculated as to what degree Tarp function contributes to Chlamydia ’s ability to parasitize mammalian host cells. To address this question, we generated a complete tarP deletion mutant using the fluorescence-reported allelic exchange mutagenesis (FRAEM) technique and complemented the mutant in trans with wild-type tarP or mutant tarP alleles engineered to harbor in-frame domain deletions.