Table 2.
Plasmids
| Plasmid | Relevant genotype | Source or reference |
|---|---|---|
| pET24a | T7 promoter, Kanr | Novagen |
| pNB100 | pBAD18-flag (3×) | 9 |
| pDSW204 | Mutant lacUV5 promoter, pBR322 origin, Ampr | 61 |
| pMER77 | Based on pDSW204 with C-terminal FLAG tag sequence | This study |
| pMER78 | Based on pNB100 with DsbA signal sequence and C-terminal FLAG tag | 58 |
| pBAD33 | Arabinose promoter, pACYC origin, Camr | 27 |
| pKD4 | pANTSγ derivative, FRTa-flanked Kan cassette from pCP15 | 17 |
| pET24a-trbB (−sp) | Mutant trbB from plasmid F with signal sequence removed | This study |
| pET24a-trbB C81S/C84S | pET24a-trbB (−sp) with C81S and C84S mutations | This study |
| pET24a-dsbC_E.coli (−sp) | Mutant dsbC from E. coli with signal sequence removed | This study |
| pDSW204-dsbC_E.coli | Wild-type dsbC from E. coli with native signal sequence | This study |
| pDSW204-dsbC_R27 | Wild-type dsbC from plasmid R27 with native signal sequence | This study |
| pDSW204-trbB | Wild-type trbB from plasmid F with native signal sequence | This study |
| pDSW204-trbB C81S/C84S | Mutant trbB from plasmid F with mutations C81S and C84S in active site | This study |
| pDSW204-trbB Δ29 | Mutant trbB from plasmid F excluding residues 21 to 50 | This study |
| pDSW204-trbB-FLAG | Wild-type trbB from plasmid F with C-terminal FLAG tag in pMER77 | This study |
| pMER78-trxA | trxA with DsbA signal sequence | 58 |
| pBAD33-appA | Wild-type appA | 58 |
| pBAD33-phoA | Wild-type phoA | 8 |
a
FRT, FLP recombination target.