FIG 4.
Direct evidence of gfp-bla excision. (A) McCoy cells infected with equal IFUs of WT, L2 tmeA, or L2Rif tmeA-lx C. trachomatis were harvested at 24 hpi, and DNA was extracted for qPCR. Relative copy numbers for tmeA, tmeB, gfp, and cre were assessed by signal normalized to chlamydial 16S rRNA. ND, none detected. (B) Excision of the reporter cassette confirmed by PCR by amplifying the tmeA locus with primers in the surrounding upstream and downstream regions. Products are shown resolved in a 1.0% agarose gel. (C) The sequenced tmeAB locus from L2Rif tmeA-lx indicating the remaining loxP scar sequence (underlined). Flanking DNA appears in blue, while start codons are in green and the tmeA stop is highlighted in red. The noncanonical start codon for TmeB is also depicted.