FIG 5.

Plasmid copy number and GlgA expression among CMUT3 transformants. HeLa cell cultures infected with wild-type (wt) C. muridarum (CM), plasmid-free (pf) CMUT3, or various CMUT3 transformants as indicated on top of the figure were harvested for quantitating genome and plasmid copies using real-time PCR, as well as various chlamydial protein levels, as listed on the right side of the figure. The ratios were calculated based on plasmid copy number per genome from each culture sample as displayed along y axis (A). Please note that although all transformants displayed significantly higher plasmid copy numbers per genome than the wt CM, there was no significant difference among the transformants. (B) Both the chlamydial major outer membrane protein (MOMP) and heat shock protein (HSP) 60 (detected by MAbs) were used as control for equal sample loading among different lanes of the Western blot. Note that although all cultures expressed similar levels of MOMP and HSP60, the CMUT3 or CMUT3 transformed with pGFPDPgp3 or DPgp4 failed to express Pgp3. More importantly, the pGFPΔPgp5 transformed CMUT3 expressed the highest level of GlgA.