TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype and/or phenotype | Source or reference |
|---|---|---|
| Strains | ||
| H. ducreyi | ||
| 35000HPa | Human-passaged variant of strain 35000 (21) | S. M. Spinola et al. (67) |
| FX517 | 35000HPdsrA::CATc | 16 |
| FX533 | 35000HPdltA::Kan | This study |
| FX534 | 35000HPdsrA::CAT dltA::Kan | This study |
| H. influenzae | ||
| Rd | Wild type | ATCCb |
| Rd/p1286 | Strain Rd containing pUNCH1286 | This study |
| Rd/pLSKS | Strain Rd containing pLSKS | This study |
| E. coli | ||
| DH5α | recA gyrB | Bethesda Research Labs |
| BL21(DE3)/pLYSs | Cmr | Invitrogen |
| Plasmids | ||
| pCR2.1-TOPO | PCR cloning vector; Kanr Ampr | Invitrogen |
| pCRT7/NT-TOPO | Expression vector; Ampr | Invitrogen |
| pLSKS | H. ducreyi shuttle vector; Smr | 77 |
| pRSM1791 | Mutagenesis plasmid; β-Gal+ Ampr | 5 |
| pUNCH1284 | dltA expression clone in pCRT7/NT-TOPO; used for rDltA purification | This study |
| pUNCH1285 | dltA PCR clone in pCR2.1-TOPO | This study |
| pUNCH1286 | dltA subclone in pLSKS | This study |
| pUNCH1287 | Mutagenized dltA; pUNCH1286 mutagenized with Kan cassette in pLSKS; Kanr Smr | This study |
| pUNCH1289 | Mutagenized dltA subclone in pRSM1791 | This study |
| pUNCH1295 | NcaA expression clone | This study |
a
35000HP was isolated from a pustular lesion of a human volunteer 13 days after inoculation.
b
ATCC, American Type Culture Collection.
c
CAT, chloramphenicol acetyltransferase.