FIG 1.

Generation of C. trachomatis ΔtarP. (A) Schematic of the tarP locus in C. trachomatis L2. FRAEM was used to delete the entire tarP (ctl0716) sequence and insert a cassette containing gfp and bla. (B) The loss of tarP and presence of gfp were confirmed in the resulting strain via qPCR. Curing of pSUmC was confirmed using primers specific for mcherry, and retention of endogenous pL2 was verified using primers specific for plasmid-encoded pgp7-8. (C) Chemiluminescence immunoblot of EB material from wild-type (WT) or mutant Chlamydia (tarp) with TarP-specific antibodies. MOMP was probed as a loading control. Cultures of HeLa cells were infected with equal IFU of WT or tarP chlamydiae. At 0, 12, 24, 36, and 48 h postinfection, one replicate was processed for enumeration of progeny EBs on fresh HeLa monolayers (D) while the other was fixed and stained for measurement of inclusion areas (24 h is shown) (E).