Table 2.
Strains and plasmids used in this study
| Strains or plasmids | Relevant property | Refs. |
|---|---|---|
| V. cholerae | ||
| 2477c | O1 El Tor, Ogawa | Lab collections stock |
| N16961 | Spontaneous mutant of N16961, Inaba, Smr | Lab collections stock |
| ICDC-VC5029 | O1 El Tor, Ogawa, with natural resistance phenotype resistant phenotype control | Lab collections |
| NΔOS | Core OS mutant strain, VC0231 deletion of N16961 (Smr) | 1 |
| NΔtolC | tolC (VC2346) deletion of N16961 (Smr) | This study |
| NΔtolC-OS | tolC and OS deletion of N16961 (Smr) | This study |
| NΔtolC-C | NΔtolC strains were complemented with pSRKTc-TolC | This study |
| NΔOS-C | NΔOS strains were complemented with pBAD33-VC0231 | This study |
| NΔtolC-OS-CtolC | NΔtolC-OS strains were complemented with pSRKTc-TolC | This study |
| NΔtolC-OS-COS ΔΔVC2436ΔVC0231C0231 | NΔtolC-OS strains were complemented with pBAD33-VC0231 | This study |
| NΔtolC-OS-C | NΔtolC-OS strains were complemented with pSRKTc-TolC and pBAD33-VC0231 | This study |
| NΔtolC-CA78D | NΔtolC strains were complemented with pSRKTc-TolCA78D | This study |
| NΔtolC-CΔ290–291 | NΔtolC strains were complemented with pSRKTc-TolCΔ290–291 | This study |
| E. coli | ||
| SM10 λpir | Km, thi thr leu tonA lacY supE recA:: RP4-2-TC::Mu λpir | 2 |
| DH5α λpir | sup E44, ΔlacU169 (ΦlacZΔM15), recA1, endA1, hsdR17, thi-1, gyrA96, relA1, λpir gyrA96, relA1, λpir | Lab collections stock |
| XL1blue | endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac | Lab collections |
| BTH101 | F−, cya-99, araD139, galE15, galK16, rpsL1 (Strr), hsdR2, mcrA1, mcrB1 | 3 |
| Plasmids | ||
| pSC123 | Suicide plasmid carrying transposon; Kanr Cmr | 4 |
| pWM91 | Suicide plasmid; oriR oriT lacZ tetAR sacB | 5 |
| pWM91-ΔtolC | pWM91 carrying upstream and downstream fragments flanking tolC | This study |
| pWM91-ΔVC0231 | pWM91 carrying upstream and downstream fragments flanking VC0231 | This study |
| pUT18C (abbreviated as pT18C) | pUT18C-derived vector, designed to create C-terminal heterologous protein fusion, Ampr | 3 |
| pKT25 (abbreviated as pT25) | lac promoter and the T25 fragment for C-terminal heterologous protein fusion. Kmr | 3 |
| pSRKTc | lac promoter and lacIq, Tetr | 6 |
| pSRKTc-TolC | pSRKTc-derived, VC2436 (TolC), Tetr | This study |
| pSRKTc-TolCA78D | pSRKTc-derived, TolCA78D, Tetr | This study |
| pSRKTc-TolCΔ290–291 | pSRKTc-derived, TolCΔ290–291, Tetr | This study |
| pBAD33-VC0231 | pBAD33-derived, VC0231, Cmr | This study |
| pT18C-TolC | pUT18C-derived, TolC23-end, Ampr | This study |
| pT25-TolC | pKT25-derived, TolC23-end, Kanr | This study |
| pT25-TolCA78D | pKT25-derived, TolCA78D, Kanr | This study |
| pT25-TolC Δ290–291 | pKT25-derived, TolCΔ290–291, Kanr | This study |
| pT25-gp44 | pKT25-derived, gp44, Kanr | This study |
| pT25-gp44(245–753) | pKT25-derived, gp44 (245–753), Kanr | This study |
| pT25-gp44(451–753) | pKT25-derived, gp44 (451–753), Kanr | This study |
| pT25-gp44(1–450) | pKT25-derived, gp44 (1–450), Kanr | This study |
| pT18C-TolCA78D | pUT18C-derived, TolCA78D, Ampr | This study |
| pT18C-TolCΔ290–291 | pUT18C-derived, TolCΔ290–291, Ampr | This study |
| pGEX-6p-1-TolC | pGEX-6p-1-derived, TolC23-end, Ampr | This study |
| pGEX-6p-1-TolCA78D | pGEX-6p-1-derived, TolCA78D, Ampr | This study |
| pGEX-6p-1-TolCΔ290–291 | pGEX-6p-1-derived, TolCΔ290–291, Ampr | This study |
| pET30a-gp44 | pET30a-derived, gp44 (245–753), Kanr | This study |
| pET30a-gp44(451–753) | pET30a-derived, gp44 (451–753), Kanr | This study |
| pET30a-gp44(1–450) | pET30a-derived, gp44 (1–450), Kanr | This study |