Table 1.
Escherichia coli strains and plasmids used in this study.
| Strain or plasmid | Genotype or main characteristics | Purpose | Origin/References |
|---|---|---|---|
| STRAINS | |||
| TG1 | supE Δ(mcrB-hsdSM)5 thi-1 Δ(lac–proAB) F'[traD36 proAB lacIq lacZΔM15] | plasmid propagation | Lucigen |
| HI-control 10G | mcrA Δ(mrr-hsdRMS-mcrBC) endA1 recA1 Φ80dlacZΔM15 ΔlacX74 araD139 Δ(ara,leu)7697 galU galK rpsL (StrR) nupG λ−tonA /Mini-F lacIq1(GentR) | plasmid propagation | Lucigen |
| BL21 (DE3) | [F−ompT, hsdS(rB−, mB−), gal, dcm λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5] | RegA-HTH-6His production | Novagen |
| HI-control BL21(DE3) | F- ompT hsdSB(rB- mB-) gal dcm λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]/mini-F lacIq1(GentR) | 6His-SUMO-RegA, 6His-SUMO-RegA_D68A and 6His-SUMO-RegA_A102S production | Lucigen |
| DH5α | [F−Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rK–, mK+) phoA supE44 λ– thi-1 gyrA96 relA1] | regA mutagenesis | Taylor et al., 1993 |
| PLASMIDS | |||
| pET21 | ampicillin resistance; T7 transcription start; C-terminal 6His•Tag sequence | RegA-HTH-6His production | Novagen |
| pETite N-His Sumo kan | kanamycin resistance; T7 transcription start; N-terminal 6His•SUMO Tag sequence | 6His-SUMO-RegA, 6His-SUMO-RegA_D68A and 6His-SUMO-RegA_A102S production | Lucigen |
| RegA-HTH-6His/pET21 | DNA binding domain of RegA fused to a C-terminal 6His•Tag sequence | RegA-HTH-6His production | This study |
| 6His-SUMO-RegA/pETite plasmid | RegA fused to a N-terminal 6His•SUMO | 6His-SUMO-RegA production | This study |
| 6His-SUMO-RegA_D68A/pETite | the conserved D68 residue of RegA was substituted for an alanine residue | 6His-SUMO-RegA_D68A production | This study |
| 6His-SUMO-RegA_A102S/pETite | the conserved A102 residue of RegA was modified to a serine residue | 6His-SUMO-RegA_A102S production | This study |