Table 1.
Bacterial strains and plasmids used in this study
| Bacterial strains/Plasmids | Description | Source |
|---|---|---|
| E. coli K12 BW25113 strains | ||
| Wild-type | rrnB3 ΔlacZ4787 hsdR514 Δ(araBAD)567 Δ(rhaBAD)568 rph-1 | (35) |
| Δrac | Whole prophage rac removed | (38) |
| Δhfq | Δhfq Δkan | This study |
| ΔralR | ΔralR Δkan | This study |
| ΔralRA | ΔralRA Δkan | This study |
| Plasmids | ||
| pCA24N | CmR; lacIq | (36) |
| pCA24N-ralR | CmR; lacIq, PT5-lac::ralR | (36) |
| pCA24N-ralA | CmR; lacIq, PT5-lac::ydac | (36) |
| pCA24N-ralR-ralA | CmR; lacIq, PT5-lac::ralR-ydaC | This study |
| pBAD-ralA | AmpR; PBAD::ralA+ | This study |
| pCA24N-hfq | CmR; lacIq, PT5-lac::hfq | (36) |
| Mutated pCA24N-ralR-ralA plasmids | This study | |
| pCA24N-ralR-ralA-G81CG83A | G to C at position 81 in ralA; G to A at position 83 in ralA | |
| pCA24N-ralR-ralA-G83A | G to A at position 83 in ralA | |
| pCA24N-ralR-ralA-G289del | Sequences removed after G at position 289 in ralR-ralA | |
| pCA24N-ralR-T189C-ralA | T to C at position 189 in ralR, changed putative ydaC start aa from Met to Thr, no change in RalR aa sequence | |
| pCA24N-ralR-ralA-C161A | C to A at position 161 in ralA, introduced a stop codon TAA at the ninth aa of putative YdaC | |
| pCA24N-ralR-ralA-G64C | G to C at position 64 in ralA | |
| pCA24N-ralR-ralA-T63C | T to C at position 63, no change in aa sequence | |
| pCA24N-ralR-ralA-M1 | Modified the 16-nt repeat in ralA to AGCCTCCTTCTTACCT, no change in aa of putative YdaC | |
| pCA24N-ralR-M2-ralA | Modified the 16-nt repeat in ralR, change aa from Gly-Ser-Glu-Lys-Glu-Ala to Gly-Ser-Ala-Ile-Ala-Glu | |
| pCA24N-ralR-M1-ralA-M1 | Modified the 16-nt repeat both in ralR and ralA to AAGTGAGAAGGAGGCT and AGCCTCCTTCTTACCT | |
| Mutated pCA24N-ralR plasmids | This study | |
| pCA24N-ralR-A154G | Encodes RalR-mutant RalR K52G | |
| pCA24N-ralR-M1 | Modified the 16-nt repeat in ralR to AAGTGAGAAGGAGGCT, no change in aa | |
| pCA24N-ralR-M2 | Modified the 16-nt repeat in ralR, change aa from Gly-Ser-Glu-Lys-Glu-Ala to Gly-Ser-Ala-Ile-Ala-Glu | |
| Note. Cm R and Amp R indicate chloramphenicol and ampicillin resistance, respectively, and aa indicates amino acid. | ||