Figure 7.

The protease domain of VgrG2b is required for its antibacterial activity. Serial dilutions (from non-diluted to 10⁻⁵) of normalized cultures of E. coli BL21(DE3)pLys producing the wild-type C-terminal domain of VgrG2b in the cytoplasm, called CterVgrG2b^C (from pBB43, a pETDuet-1 derivative, line 5) or in the periplasm, called CterVgrG2b^P (from pBB44, a pET22b(+) derivative yielding a fusion of the C-terminal domain with a Sec signal peptide, line 2) or two variants CterVgr2b^P_H935A (from pBB45, line 3) and CterVgr2b^P_E936A (from pBB46, line 4) were spotted on LB agar plates supplemented (left panel) with 0.4% glucose or (right panel) with 0.1 mM IPTG. In line 6, the CterVgrG2b^P (from pBB44) was coproduced with Tli3 (from pVT8) and in line 7, the CterVgrG2b^P and Tli3 were coproduced from the same transcript (from pSBC110). Glucose and IPTG allow respectively repression and induction of the gene encoding the T7 RNA polymerase.