FIG 6.

Attenuation of C. trachomatis tarP in a murine model can be reversed by cis complementation with WT tarP. (A) Schematic of allelic replacement strategy to restore tarP to the mutant strain. FRAEM was used to replace the gfp-bla cassette in the ΔtarP mutant with the tarP gene and a downstream aadA selection marker. (B) Representative fluorescence-based Western blot of tarP levels in 24-h culture material derived from equal infections with wild-type (WT), mutant (tarp), and cis-complemented (cis-tarp) Chlamydia. Hsp60 was probed as a loading control. (C) Signal intensity of Tarp-specific signal was normalized to Hsp60, and values are plotted as a function WT Tarp signal. Error bars represent one standard deviation from blots derived from 3 separate experiments. Groups of 5 female C3H/HeJ mice were infected intravaginally with equal input IFUs, and shed IFUs were enumerated every 4 days from day 3 to 31 postinfection. Shed bacteria were enumerated by passage in HeLa cells and are represented as number of mice actively shedding detectible chlamydiae (D) or numbers of detected inclusions (E). Data are represented with standard deviation, and two-way repeated measures (RM) ANOVA was performed to establish statistical significance (*, P < 0.0001) compared to the wild type.