Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or descriptiona | Source or reference |
|---|---|---|
| E. coli | ||
| TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu)7697 galU galK rpsL (strR) endA1 nupG | Invitrogen |
| HB101 | F thi-1 hsdS20(rB− mB−) supE44 recA13 ara-14 leuB6 proA2 lacY1 galK2 rpsL20 (strR) xyl-5 mtl-1 | Promega |
| DY380 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 deoR recA1 endA1 araD139 Δ(ara leu)7649 galU galK rspL nupG [λ cI857 Δ(cro-bioA) (tetR)] | 50 |
| MEB149 | DY380 with plasmid pMEB144 | This study |
| H. ducreyi | ||
| 35000HP | Class I clinical isolate; human-passaged variant of strain 35000; Winnipeg, Canada, 1975 | 3 |
| HD183 | Class I clinical isolate; Singapore, 1982 | 9, 49 |
| HD188 | Class I clinical isolate; Kenya, 1982 | 9, 49 |
| 82-029362 | Class I clinical isolate; California, 1982 | 9, 49 |
| 6644 | Class I clinical isolate; Massachusetts, 1989 | 9, 49 |
| 85-023233 | Class I clinical isolate; New York, 1985 | 9, 49 |
| CIP542 ATCC | Class II clinical isolate; Hanoi, Viet Nam, 1954 | 23, 54 |
| HMC112 | Class II clinical isolate; 1984 (location not reported) | 54 |
| 33921 | Class II clinical isolate; Kenya (yr not reported) | 9, 15 |
| DMC164 | Class II clinical isolate; Bangladesh (yr not reported) | 54 |
| 35000HP(pLSSK) | 35000HP with vector pLSSK; Strr | This study |
| 35000HPmtrC | mtrC unmarked deletion mutant of 35000HP | This study |
| 35000HPmtrC(pLSSK) | 35000HPmtrC with vector pLSSK; Strr | This study |
| 35000HPmtrC(pmtrC) | mtrC unmarked deletion mutant of 35000HP with plasmid pMEB219; Strr | This study |
| FX517 | dsrA::cat insertion mutant of 35000; Cmr | 17 |
| 35000HPΔcpxA | cpxA unmarked deletion mutant of 35000HP | 48 |
| 35000HPΔcpxR | cpxR::cat insertion/deletion mutant of 35000HP; Cmr | 28 |
| Plasmids | ||
| pGEM-T Easy | TA cloning vector; Ampr | Promega |
| pCR-XL-TOPO | TA cloning vector; Kanr | Invitrogen |
| pRSM2832 | Specr cassette flanked by FTR sites | 50 |
| pRSM2975 | FLP recombinase vector | 48 |
| pLSSK | H. ducreyi shuttle vector; Strr | 55 |
| pRSM2072 | Suicide vector; Ampr | 12 |
| pMEB144 | mtrC + flank in pGEM-T Easy | This study |
| pMEB191 | mtrC replaced with Specr cassette in pGEM-T Easy | This study |
| pMEB193 | mtrC replaced with Specr cassette in pRSM2072 | This study |
| pMEB219 | mtrC in pLSSK | This study |
a
Strr, resistance to streptomycin; Cmr, resistance to chloramphenicol; Ampr, resistance to ampicillin; Kanr, resistance to kanamycin; Specr, resistance to spectinomycin.