TABLE 4.
Plasmids and strains used in this study
| Name of plasmid or strain | Relevant featuresa | Source or reference |
|---|---|---|
| Plasmids | ||
| pAP24 | Ptet-slucopt; catP | 24 |
| pCD22 | PtcdA; catP | 53 |
| pIB14 | PT7-sigB6×His; amp | This study |
| pIB21 | Pcd0872-slucopt; catP | This study |
| pIB27 | Pcd3412-slucopt; catP | 20 |
| pIB68 | Pcd0350-slucopt; catP | 20 |
| pIB69 | Pcd2963-slucopt; catP | 20 |
| pIB73 | Pcd3605-slucopt; catP | This study |
| pIB74 | Pcd3614-slucopt; catP | 20 |
| pMTL007C-E2_sigB171s::intron_ermB | sigB-retargeted intron (ermB) | This study |
| pWKS1750 | Ptet-sigB; catP | This study |
| Strains | ||
| 630Δerm | MLS-susceptible derivative of Clostridioides difficile strain 630 | 54, 55 |
| AP34 | 630Δerm pAP24; Thiar | 24 |
| DH5α | Escherichia coli F− endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG purB20 ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 hsdR17(rK− mK+) λ− | Laboratory stock |
| IB14 | Rosetta (DE3) pLysS pIB14; Ampr, Chlorr | This study |
| IB18 | 630Δerm sigB::CT; Ermr/Lincor | This study |
| IB37 | 630Δerm pIB27; Thiar | 20 |
| IB56 | 630Δerm ΔsigB | 20 |
| IB58 | IB18 pWKS1750; Thiar, Ermr/Lincor | This study |
| IB61 | IB18 pAP24; Thiar, Ermr/Lincor | This study |
| IB95 | 630Δerm pIB68; Thiar | 20 |
| IB96 | 630Δerm pIB69; Thiar | 20 |
| IB98 | IB56 pIB27; Thiar | 20 |
| IB99 | IB56 pIB68; Thiar | 20 |
| IB100 | IB56 pIB69; Thiar | 20 |
| IB108 | 630Δerm pIB74; Thiar | 20 |
| IB111 | IB56 pIB74; Thiar | 20 |
| JC096 | 630Δerm pWKS1750; Thiar | This study |
a
MLS, macrolides-lincosamides-streptogramin B; Amp, ampicillin; Chlor, chloramphenicol; Erm, erythromycin; Linco, lincomycin; Thia, thiamphenicol; r, resistance.