Figure 1.

Over expression of RelA-C-terminus domain (CTD) on cell growth. (A) RelA domains as described in Loveland et al. (2016) and YG4 fragment. (B,C) W3110 [wild type (WT)] cells bearing a plasmid overexpressing YG4 or YG4-C638F were grown in Luria-Bertani (LB) medium for 2 h, after which overexpression was induced by the addition of 1 mg/ml of IPTG for 1 h. Cells were collected and washed, diluted in serial dilutions, and plated on M9 medium. ΔRelA and WT + pQE (an empty plasmid) were used as controls. (B) containing AT; (C) without AT. All plates were incubated at 37°C overnight. WT cells with an “empty” plasmid (WT) and cells deleted for RelA (ΔRelA) were used as controls. (D) All cell types were grown in duplicates in a 24-well plate in LB medium supplemented with 100 μg/ml of ampicillin. After 2 h of growth, all cells were supplemented with 1 mg/ml of IPTG and were grown for an additional 2.5 h at 37°C with shaking. Cell growth was monitored by optical density (OD) measuring OD₆₀₀. No bullets – WT cells with an empty plasmid; square (▪) – WT cells overexpressing YG4; triangle (▲) – WT cells overexpressing Rel251; dashed – WT cells overexpressing Rel-C638F; dots – WT cells overexpressing RelA.