Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or relevant characteristics | Reference or source |
|---|---|---|
| Strains | ||
| DH5α | F− (ϕ80lacZΔM15) Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 (rK− mK+) phoA supE44 thi-1 gyrA96 relA1 | Laboratory collection |
| Rosetta (DE3) | lon and ompT protease-deficient E. coli carrying pRARE; expresses six rare tRNAs which facilitate expression of genes that contain rare E. coli codons; Camr; p15a ori | Merck KGaA, Darmstadt, Germany |
| SPC105 | MC4100 (ΔlacU169 araD rpsL relA thi flbB) containing a chromosomal PmarII::lacZ fusion at the λ attachment site; wild-type mar locus | 6 |
| SPC107 | SPC105 with a 39-kb deletion that includes the mar locus | 23 |
| AG100 | argE3 thi-1 rpsL xyl mtl supE44 λ lysogen | 24 |
| AG112 | AG100 marR mutant | 25 |
| Plasmids | ||
| pGEM-T Easy | Vector for direct cloning of PCR fragments; Ampr | Promega, Madison, WI |
| pET28a | Expression cloning vector; Kanr; the transcription of the cloned gene is driven by the T7 RNA polymerase and controlled by the LacI repressor; ori ColE1 | Merck KGaA |
| pET21b | Expression cloning vector; Ampr; the transcription of the cloned gene is driven by the T7 RNA polymerase and controlled by the LacI repressor; ori ColE1 | Merck KGaA |
| pACT7Sp | T7 RNA polymerase regulated by lacUV5; Spr; this plasmid is used in conjunction with pET28a; ori p15a | 26 |
| pET13a-G95S | pET13a carrying G95S variant of marR; Kanr | 22 |
| pET13a-D26N | pET13a carrying D26N variant of marR; Kanr | 22 |
| pVDMarR | pET28a carrying wild-type marR; Kanr | This study |
| pVD57 | pET28a carrying P57A variant of marR; Kanr | This study |
| pVD72 | pET28a carrying T72A variant of marR; Kanr | This study |
| pVD86 | pET28a carrying R86A variant of marR; Kanr | This study |
| pVD74 | pET28a carrying M74A variant of marR; Kanr | This study |
| pVD77 | pET28a carrying R77A variant of marR; Kanr | This study |
| pVD5772 | pET28a carrying P57A-T72A variant of marR; Kanr | This study |
| pVD7477 | pET28a carrying M74A-R77A variant of marR; Kanr | This study |
| pVD26 | pET28a carrying D26N variant of marR; Kanr | This study |
| pVD95 | pET28a carrying G95S variant of marR; Kanr | This study |
| pVD16 | pET28a carrying R16A variant of marR; Kanr | This study |
| pET28a-MarR-H19A | pET28a carrying H19A variant of marR; Kanr | This study |
| pET28a-MarR-K44A | pET28a carrying K44A variant of marR; Kanr | This study |
| pVDOR | pET21b carrying ompR nucleotide sequence cloned in frame with a polyhistidine tag at the C-terminal end | This study |