TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | Host strain for cloning experiments | 51 |
| HB101 | Host strain essential for propagating plasmids carrying mutated H. ducreyi DNA inserts for use in electroporation | 51 |
| χ705 | F−leu-4 φr Strrarg-35 T6r λ− | 14 |
| χ711 | Deletion mutant of χ705 lacking the gmhA and proAB genes; expresses a truncated LPS molecule which migrates very rapidly in SDS-PAGE relative to that of χ705 | 7, 14 |
| H. ducreyi | ||
| 35000 | Wild-type strain isolated in Winnipeg, Manitoba, Canada; expresses a LOS that 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 | 60 |
| 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 | This study |
| Plasmids | ||
| pWKS30 | Low-copy-number cloning vector; Ampr | 67 |
| pHD250 | pWKS30 with an 18-kb H. ducreyi 35000 DNA insert containing the gmhA gene | This study |
| pBluescript II KS+ (pBS) | Cloning vector; Ampr | Stratagene |
| pHD251 | pBS with a 1.6-kb PCR-derived insert containing the H. ducreyi 35000 gmhA gene | This study |
| pHD252 | pHD251 with a cat cartridge inserted into the Ppu10I site within the gmhA gene | This study |
| pCR2.1 | Cloning vector; Ampr | Invitrogen |
| pCR253 | pCR2.1 with a 1-kb PCR-derived insert containing the H. ducreyi gmhA gene | This study |
| pLS88 | Cloning vector capable of replication in H. ducreyi; Kanr Smr Sulr | 16 |
| pLS253 | pLS88 with a 1-kb PCR-derived insert containing the H. ducreyi gmhA gene | This study |