TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or descriptiona | Source or reference |
|---|---|---|
| Strains | ||
| E. coli strains | ||
| DH5αλpir | F−endA1 hsdR17 supE44 thi-1 recA1 gyrA96 relA1 Δ(lacZYA-argF)U169 φ80dlacZΔM15 λpir | Laboratory collection |
| DH10B | F−mcrA Δ(mrr hsdRMS mcrBC) φ80dlacZΔM15 ΔlacX74 endA1 recA1 deoR Δ(ara leu)7697 araD139 galU galK nupG rpsL | Laboratory collection |
| SM10λpir | thi thr leu tonA lacY supE recA::RP4-2-Tc::Mu λpir pro endA hsdA hsdR supF | Laboratory collection |
| V. cholerae strains | ||
| N16961 | Wild-type El Tor strain, Smr | 10 |
| LK626 | N16961 ΔVC2773 | This study |
| LK681 | N16961/pJK4 | This study |
| LK705 | N16961 ΔVC2061 | This study |
| LK706 | N16961 ΔVC2773 ΔVC2061 | This study |
| LK712 | LK626 with reconstituted VC2773 | This study |
| KFV10 | N16961 ΔlacZ | 7 |
| Plasmids | ||
| pBluescript SKII+ | Cloning vector, Ampr | Stratagene |
| pCVD442 | oriR6K mobRP4 bla sacB | 4 |
| pWM91 | oriR6K oriTRP4bla sacB | 22 |
| pRKlac290 | Broad-host-range vector with promoterless lacZ | 28 |
| pWKS30-Tc | Low-copy-number broad-host-range vector, Tcr | 14 |
| pJK4 | pWKS30-Tc with parAB upstream region and parA | This study |
| pSD15 | VC2773 in-frame deletion construct in pCVD442 | This study |
| pJK11 | VC2061 in-frame deletion construct in pWM91 | This study |
| pSD19 | pRKlac290 with 450-bp parAB upstream region | This study |
| pDAG114 | Stable mini-F, Cmr | 25 |
| pDAG203 | Unstable mini-F, Δ(sopOPABC) Cmr | 5 |
| pSD38 | pDAG203 with IR1 cloned into AflIII-Bsu36I sites | This study |
| pSD39 | pDAG203 with IR2 cloned into AflIII-Bsu36I sites | This study |
| pSD44 | pDAG203 with IR3 cloned into AflIII-Bsu36I sites | This study |
| pSD25 | pBluescript II SK(+) with parAB and upstream region | This study |
| pSD9 | pBluescript II SK(+) with parB and in-frame deletion of parA | This study |
| pSD21 | pBluescript II SK(+) with parA and in-frame deletion of parB | This study |
a
Ampr, ampicillin resistance; Tcr, tetracycline resistance; Cmr, chloramphenicol resistance; Smr, streptomycin resistance.