TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| V. cholerae strains | ||
| O395 | Wild type, Strr | Laboratory strain |
| RY1 | O395 ΔtcpP | 26 |
| RY6 | O395 toxTΔpro | 26 |
| EK307 | O395 ΔtoxR | 11 |
| EK459 | O395 ΔtoxR ΔtcpP | 11 |
| EK1647 | O395 ΔtcpP toxTΔpro | This study |
| EK1654 | O395 ΔtoxR ΔtcpP toxTΔpro | This study |
| TG128 | EK459/pBluescript SK(+)/pMMB207 | This study |
| TG129 | EK459/pSK-toxR-HA/pMMB207 | This study |
| E. coli strains | ||
| DH5α | supE44 ΔlacU169 (F80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Laboratory collection |
| DH5α λpir | supE44 ΔlacU169 (F80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 (λpir) | Laboratory collection |
| SM10 λpir | thi-1 thr leu tonA lacY supE recA::RP4-2-Tc::Mu Kmr (λpir) | Laboratory collection |
| TG389 | DH5α/pREP4/pBluescript KS(−) | This study |
| TG387 | DH5α/pREP4/pTG64 | This study |
| Plasmids | ||
| pBluescript SK(+) | Apr; high-copy-no. cloning vector | Stratagene |
| pBluescript KS(−) | Apr; high-copy-no. cloning vector | Stratagene |
| pSK-toxR-HA | pBluescript SK(+)-toxR-HA | this study |
| pMMB207 | IPTG-inducible expression vector; CmrlacIqtacP | 19 |
| pTL61T | Apr; promoterless lacZ fusion vector | 13 |
| pKAS32 | Suicide plasmid; Apr Sms | 24 |
| pKAS32-tcpF-toxTΔpro-toxT | For assembly of the parental toxT promoter | 26 |
| pKAS32-tcpF-toxTpro-toxT | For introducing transversion mutations onto the chromosome | This study |
| pEK41 | pMMB207-tcpP-HSV | 11 |
| pREP4 | p15A, lacIq Kmr | Qiagen |
| pRK2013 | ColE1::RK2 tra+ Kmr | 3 |
| pTG24 | Modified form of pTL61T with NotI site inserted into the XhoI, XhoI site disrupted | This study |
| pTG49 | pBluescript SK(+) with tacPtcpP-HSV inserted as a PstI/EcoRV fragment | This study |
| pTG64 | pBluescript KS(−)-tacP-tcpP-HSV | This study |
| pBluescript SK(+)-toxTpro | Vector for generation of transversion mutations and cloning into pTL61T | This study |
| pBluescript SK(+)-tcpF-toxTpro-toxT | Vector for generation of transversion mutations and cloning into pKAS32 | This study |