TABLE 3.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics or usea | Reference or source |
|---|---|---|
| C. diphtheriae strains | ||
| 1737 | Wild type, Gravis biotype, tox+ | 67 |
| 1737ΔhtaA | Deletion of htaA in 1737 | 24 |
| 1737ΔchtA | Deletion of chtA in 1737 | 44 |
| 1737ΔchtC | Deletion of chtC in 1737 | 44 |
| 1737ΔchtA/ΔchtC/ΔhtaA | Deletion of chtA, chtC, and htaA in 1737 | 44 |
| 1737ΔchtA/ΔchtC | Deletion of chtA and chtC in 1737 | 44 |
| 1737ΔhbpA | Deletion of hbpA in 1737 | This study |
| 1737Δ4 | Deletion of chtA, chtC, htaA, and hbpA in 1737 | This study |
| 1737ΔdtxR | Deletion of dtxR in 1737 | This study |
| E. coli strains | ||
| BL21(DE3) | Protein expression | Novagen |
| DH5α | Cloning strain | Invitrogen |
| S17-1 | RP4 mobilization functions | 61 |
| EPI400 | Cloning strain | Lucigen |
| One Shot TOP10 | Cloning of PCR fragments | Invitrogen |
| Plasmids | ||
| pET24(a)+ | Expression vector, Knr | Millipore |
| pET-hbpA | Strep-tag coding sequence-tagged hbpA cloned into pET24(a)+ | This study |
| pKN2.6Z | C. diphtheriae shuttle vector, Knr | 41 |
| pET-htaA | Strep-tag coding sequence-tagged htaA cloned into pET24(A)+ | 42 |
| pET-Cterm | Strep-tag-tagged C-terminal domain of ChtA in pET24(a)+ | 44 |
| pKN-htaA | pKN2.6Z carrying the htaA gene | 24 |
| pKN-chtC | pKN2.6Z carrying the chtC gene | 43 |
| pKN-hbpA | pKN2.6Z carrying the hbpA gene | This study |
| pK18mobsacB | C. diphtheriae shuttle vector, Knr | 62 |
| pSPZ | Promoter probe vector, Spcr | 60 |
| pSPZ-hbpA | pSPZ with hbpA promoter region | This study |
| pCR-Blunt-II-TOPO | Cloning of PCR fragments, Knr | Invitrogen |
| pMS298 | Encodes C. diphtheriae dtxR under T7 control, Ampr | 63 |
| pGP1-2 | Encodes temp-inducible T7 RNA polymerase, Knr | 71 |
a
Kn, kanamycin; Spc, spectinomycin; Amp, ampicillin.