TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Description or sequence |
|---|---|
| Strains | |
| C7258 | Wild type, El Tor biotype (Perú isolate, 1991) |
| WL7258 | C7258 Δcrp (15) |
| AJB51 | C7258 ΔhapR (15) |
| WL51 | C7258 ΔhapR Δcrp (15) |
| WL52 | C7258 ΔhapR Δcya (this study) |
| Plasmids | |
| pTTVpsR | 1.4-kb BamHI-EcoRI DNA fragment encoding vpsR ORF in pTTQ18 |
| pBADCRP7 | crp ORF cloned in pBADHisB (23) |
| pCya1 | 0.9-kb EcoRI-BamHI DNA fragment 5′ of cya ORF in pUC19 |
| pCya2 | 0.8-kb BamHI-XbaI DNA fragment 3′ of cya ORF in pUC19 |
| pΔCya | EcoRI-BamHI and BamHI-XbaI fragments from pCya1 and pCya2 in pUC19 |
| pΔCya-Km | 1.2-kb BamHI Kmr cassette from pUC4K cloned in pΔCya |
| pCVDΔCya-Km | 2.9-kb EcoRI/Klenow-XbaI fragment from pΔCya-Km in pCVD442 |
| pTT-Cya | Cya ORF in pTTQ18 |
| Primers | |
| CytR295 | 5′-CAACAGAAGCGTCGGGAGAACTC |
| CytR421 | 5′-CGCAAATTCACACGCCATCACCAT |
| Cya7 | 5′-CGGAATTCCTTGCAGGCTTATACTCAG |
| Cya902 | 5′-CGGGATCCAGTCCATGCCGTACAGAT |
| Cya1759 | 5′-CGGGATCCTGGTGCTGAGCAGAAATG |
| Cya2567 | 5′-GCTCTAGAGATTTAGAAAACCTGAGACG |
| RecA578 | 5′-GTGCTGTGGATGTCATCGTTGTTG |
| RecA863 | 5′-CCACCACTTCTTCGCCTTCTTTGA |
| VpsA434 | 5′-ACCACTTTGCACCTACAGATACTTC |
| VpsA676 | 5′-CGGTAGTGATCAGCGCTTGGCAA |
| VpsL607 | 5′-ACTGGGCAGGTGCAAAATGTCTATA |
| VpsL775 | 5′-AGGGGGTATCAAAAATGCTAAACGC |
| VpsR75 | 5′-GGCTGTGTTGGAAAAAGTGGGTTG |
| VpsR206 | 5′-GGCTACTCACCAAATTCGCAATCC |
| VpsR78 | 5′-GCGAATTCCATGAGCACTCAATTCCGTA |
| VpsR1431 | 5′-GCGGATCCCAAGGTAAATCAGCAAAAC |
| VpsT56 | 5′-CCAGATTGTTGAAAGAGGCGTTAG |
| VpsT252 | 5′-TGCGGACAGTTTATGATGACCTCT |