A schematic of constructed plasmids using CT695 as an example.

<p>(A). pUCNmP. The <i>Neisseria meningitidis</i> promoter (NmP) was inserted into pUC19 upstream from BlaM. Insertion/Deletion PCR is used to insert any chlamydial sequence (<i>ct695</i> is shown) to create a translational fusion of the chlamydial gene (green) with the β-lactamase gene (blue). DNA elements can then be PCR amplified using primers NmP+BlaFus+AscI F and NmP+BlaFus+AscI R to generate a product flanked by AscI restriction sites. (B). pL2dest was created by replacement of the coding sequence for GFP/CAT of pGFP::SW2 with the mCherry gene. A chloramphenicol drug cassette flanked by AscI recognition sequences was introduced immediately downstream from mCherry coding sequence. (C). pCT659-BLA was created by ligation of AscI-digested PCR product into the AscI site in pL2dest. The resulting plasmid allows expression of CT695-Bla from the constitutive Nmp promoter.