TABLE 1.
Plasmids used in this study
| Plasmid | Relevant details | Ref. or source |
|---|---|---|
| pCBR023 | Pcwp2 driving codon-optimized gusA gene in pUC19 | Ref. 15 |
| pRPF137 | EcoRI site between Pcwp2 and gusA in pCBR023 changed to SacI | This study |
| pMTL960 | E. coli-C. difficile shuttle vector | Nigel Minton |
| pRPF144 | Pcwp2-gusA cassette from pRPF137 subcloned into pMTL960 | This study |
| pRPF150 | gusA gene in pRPF144 replaced with WT secA2 with N-terminal Strep-tag II | This study |
| pRPF151 | gusA gene in pRPF144 replaced with secA2(K106R) with N-terminal Strep-tag II | This study |
| pSTBlue-1 | E. coli cloning vector | Novagen |
| pRMC2 | Tetracycline-inducible vector for S. aureus | Ref. 36 |
| pRPF135 | Ptet from pRMC2 cloned into pSTBlue-1 | This study |
| pRPF139 | Extraneous BamHI and XhoI sites in pRPF135 removed | This study |
| pRPF141 | Extraneous KpnI site in pRPF139 removed | This study |
| pRPF143 | Pcwp2 in pRPF137 replaced with modified Ptet from pRPF141 | This study |
| pRFP146 | Ptet-gusA cassette from pRPF143 cloned into pMTL960 | This study |
| pRPF177 | pRPF146 with slpA transcriptional terminator added after gusA | This study |
| pRPF185 | pRPF177 with fdx transcriptional terminator added after tetR | This study |
| pRPF186 | WT secA2 with N-terminal Strep-tag II cloned into pRPF185 | This study |
| pRPF187 | secA2(K106R) with N-terminal Strep-tag II cloned into pRPF185 | This study |
| pRPF193 | WT secA1 with N-terminal Strep-tag II cloned into pRPF185 | This study |
| pRPF194 | secA1(K106R) with N-terminal Strep-tag II cloned into pRPF185 | This study |
| pRPF195 | Inducible antisense RNA to secA2 | This study |
| pRPF204 | Inducible antisense RNA to secA1 | This study |