TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| Bacterial strains | ||
| E. coli | ||
| DH5α | Host strain for cloning experiments | 59 |
| HB101 | Host strain essential for propagating plasmids carrying mutated H. ducreyi DNA inserts used in electroporation | 6, 59 |
| S. typhimurium | ||
| SL3770 | Expresses wild-type LPS | 58 |
| SL3789 | rfaF (waaF) mutant; expresses a truncated LPS molecule which migrates rapidly in SDS-PAGE relative to SL3770 LPS | 58 |
| H. ducreyi | ||
| 35000 | Wild-type strain isolated in Winnipeg, Manitoba, Canada; LOS binds MAb 3E6 | 23 |
| 35000.7 | Isogenic LOS mutant with a cat cartridge inserted into the rfaK (lbgB) gene; expresses a truncated LOS molecule that does not bind MAb 3E6 | 68 |
| 35000.252 | Isogenic LOS mutant with a cat cartridge inserted into the Ppu10I site within the gmhA gene; expresses a truncated LOS molecule that does not bind MAb 3E6 | 6 |
| 35000.10 | Isogenic LOS mutant with an Ω-Cm cartridge inserted into the MscI site within the waaF gene; expresses a truncated LOS molecule that does not bind MAb 3E6 | This study |
| Plasmids | ||
| pUC19 | Cloning vector; encodes Ampr | 59 |
| pHD1.3 | pUC19 containing a 1.5-kb insert of H. ducreyi 35000 chromosomal DNA with an incomplete waaF ORF | This study |
| pBR322 | Cloning vector; encodes Ampr, Tetr | 59 |
| pSLR1 | pBR322 with a 5.8-kb H. ducreyi 35000 DNA insert containing the waaF gene | This study |
| pBB10 | pSLR1 with an Ω-Cm cartridge inserted into the MscI site within the waaF gene | This study |
| pCR2.1 | Cloning vector; encodes Ampr | Invitrogen |
| pBB12 | pCR2.1 with a 1.5-kb PCR-derived insert containing the H. ducreyi waaF gene | This study |
| pLS88 | Cloning vector capable of replication in H. ducreyi; encodes Kanr, Smr, Sulr | 15 |
| pBB13 | pLS88 with a 1.5-kb PCR-derived insert containing the H. ducreyi waaF gene | This study |