Fig 2.

Effect of pORF deletion on expression of other genes. HeLa cell were infected with wild-type L2 (L2 Wt), plasmid-free L2 [L2R(25667)], or various stable L2R transformants, including L2RpGFP::SW2, L2RpGFPΔpgp3, L2RpGFPΔpgp4, L2RpGFPΔpgp5, and L2RpGFPΔpgp7, and the infected cultures were harvested at 30 h postinfection for total RNA preparation. The transcript copy number was determined from three replicates by using gene-specific primer/probe sets and TaqMan quantitative RT-PCR and was normalized to the lpdA mRNA copy number in the corresponding samples, since lpdA is constitutively expressed. Three groups of genes were detected, including plasmid-borne genes (pgp1 to pgp8), plasmid-regulated chromosomal genes (CTL0397, CTL0814, and CTL0167 [glgA]), and the non-plasmid-regulated chromosomal gene CTL0050 (ompA) (encoding the major outer membrane protein). Note that there is a significant reduction in the expression level of plasmid-regulated genes (except for CTL0814) by L2R, L2RpGFPΔPgp3, and L2RpGFPΔPgp4 but not by L2RpGFPΔPgp5 and L2RpGFPΔPgp7 organisms.