FIG. 5.

PvdN_V5H6 is recovered in the spheroplast fraction in both wild-type and tat-deficient strains. Wild-type (WT) and tatABC (tat) P. aeruginosa mutant strains expressing pvdN_V5H6 were grown in LB medium. pvdN_V5H6 expression was induced in the exponential phase by 0.1 mM IPTG for 1 h. Cells were further collected and treated for spheroplast preparation. Wild-type and tatABC spheroplasts were further centrifuged for 10 min at 8,000 RPM. The pellet corresponds to the spheroplast (S), whereas the supernatant corresponds to periplasm (P). Samples equivalent to 0.1 OD₆₀₀ unit of original cultures were further analyzed by 11% SDS-PAGE, followed by Western blotting using anti-V5 and anti-β-lactamase antibodies. Molecular mass markers (kDa) are shown on the right.