All E. coli strains used are listed in Table 2 . Standard genetic methods including transformation and P1 vir transduction were used for strain construction.
Cells were grown in Luria-Bertani (LB; 1% tryptone, 0.5% yeast extract, 0.5% NaCl) medium at 30°C for ts strains under permissive conditions and 37°C (ftsA12) or 42°C (all others) under non-permissive conditions. All non-ts strains were grown at 37°C. Antibiotic concentrations were as previously described, and culture growth was monitored by optical density as previously reported (Haeusser et al., 2014 (link)) Detailed procedures in preparation for microscopic imaging are provided below.
Expression of cloned genes from vectors derived from pET11a (Novagen – EMD Millipore) was induced with 1 mM isopropyl-β-D-galactopyranoside (IPTG) (Fisher Scientific) and from pDSW-derived vectors with 0.5 mM IPTG. Gene expression from pKG-derived vectors (J.S. Parkinson, University of Utah) was induced with 0.1, 1.0, or 5.0 μM sodium salicylate (Mallinkrodt), as indicated in text and figure legends.
General DNA and protein manipulation and analysis, including spot titers, were performed as previously described (Haeusser et al., 2014 (link)).
Cells were grown in Luria-Bertani (LB; 1% tryptone, 0.5% yeast extract, 0.5% NaCl) medium at 30°C for ts strains under permissive conditions and 37°C (ftsA12) or 42°C (all others) under non-permissive conditions. All non-ts strains were grown at 37°C. Antibiotic concentrations were as previously described, and culture growth was monitored by optical density as previously reported (Haeusser et al., 2014 (link)) Detailed procedures in preparation for microscopic imaging are provided below.
Expression of cloned genes from vectors derived from pET11a (Novagen – EMD Millipore) was induced with 1 mM isopropyl-β-D-galactopyranoside (IPTG) (Fisher Scientific) and from pDSW-derived vectors with 0.5 mM IPTG. Gene expression from pKG-derived vectors (J.S. Parkinson, University of Utah) was induced with 0.1, 1.0, or 5.0 μM sodium salicylate (Mallinkrodt), as indicated in text and figure legends.
General DNA and protein manipulation and analysis, including spot titers, were performed as previously described (Haeusser et al., 2014 (link)).
