TABLE 2.
Strains and plasmids used in this study
| Strain or plasmid | Description or use | Reference or source |
|---|---|---|
| C. diphtheriae strains | ||
| 1737 | Wild type, Gravis biotype, Tox+ | 70 |
| 1737 ΔdtxR | Deletion of dtxR in 1737 | 71 |
| 1737 Δzur | Deletion of zur in 1737 | This study |
| 1737 ΔripA | Deletion of ripA in 1737 | This study |
| 1737 Δiut | Deletion of iut gene cluster in 1737 | This study |
| 1737 Δiut Δmnt | Deletion of iut and mnt in 1737 | This study |
| 1737 Δiut Δmnt Δtro | Deletion of iut, mnt, and tro in 1737 | This study |
| E. coli strains | ||
| DH5α | Cloning and protein expression | 72 |
| BL21(DE3) | Protein expression | 73 |
| S17-1 λpir | Mating strain | 74 |
| Plasmids | ||
| pGP1-2 | Encodes temp-inducible T7 RNA polymerase; Kanr | 75 |
| pMS298 | Encodes C. diphtheriae dtxR under T7 control; Ampr | 42 |
| pET30zur | C. diphtheriae zur cloned into pET30a | This study |
| pET30ripA | C. diphtheriae ripA cloned into pET30a | This study |
| pIutAHis | C. diphtheriae iutA cloned into pET30a | This study |
| pIutAStrepII | Strep-tagged iutA cloned into pET51b | This study |
| pIutEHis | C. diphtheriae iutE cloned into pET30a | This study |
| pIutEStrepII | Strep-tagged iutE cloned into pET51b | This study |
| pKΔiut | Suicide vector for deletion of iut gene cluster (dip0169–dip0173) | This study |
| pKΔripA | Suicide vector for deletion of ripA (dip0922) | This study |
| pKΔtro | Suicide vector for deletion of tro operon (dip0438–dip0441) | This study |
| pKΔmnt | Suicide vector for deletion of mnt operon (dip0615–dip0618) | This study |
| pK19mobsacBΔzur | Suicide vector for deletion of zur (dip1710) | 33 |
| pSPZ | Carries lacZ; Spcr | 76 |
| pSPZiutA | iutA-lacZ promoter fusion | This study |
| pSPZiutE | iutE-lacZ promoter fusion | This study |
| pSPZiutE(FL) | iutE(FL)-lacZ promoter fusion | This study |
| pSPZiutE5′ | iutE5′-lacZ promoter fusion | This study |