Fig 8.

Expression of the rcc00555 endolysin and rcc00556 holin in E. coli. The lytic and peptidoglycan-degrading activities of rcc00555 (A and B) and holin-like activity of rcc00556 (C and D) expressed in E. coli are shown. (A) Turbidity after resuspension in dH₂O of induced E. coli. (B) Lane 1, Zymogram of E. coli culture lysate; lane 2, affinity-purified 555C protein; lane 3, SDS-PAGE of purified 555C protein. The approximate migration of the markers is indicated. (C) Growth curves of E. coli. Arrow, induction time point. (D) Propidium iodide staining of E. coli after induction of rcc00556 expression. Cells contained the inducible rcc00555 expression plasmids pET-555C (C-terminally 6-His tagged) or the empty vector control pET28a(+) (A and B) and the inducible rcc00556 expression plasmid pI556 or the empty vector control pIND4 (C and D). Cells were induced by addition of 1 mM IPTG. Error bars represent the ranges for two (A) or the standard deviations for three (C) biological replicates.