TABLE 1.
Bacterial strains and plasmids used in this study
| Strain | Genotype/descriptiona | Reference |
|---|---|---|
| V. cholerae | ||
| MD 992 | AM19226 R− M+ Strr | Laboratory stock |
| MD 996 | AM19226 R− M+ ΔlacZ Strr | Laboratory stock |
| AAC40 | MD992 ΔvttRB (A33_1675) | This study |
| AAC228 | MD992 ΔvttRA (A33_1664) | This study |
| MD1069 | MD992 ΔvttRA (A33_1664) ΔvttRB (A33_1675) | This study |
| AD10 | MD992 ΔtoxR | This study |
| AAC66 | MD 996(pAAC3-A33_1675-lacZY) | This study |
| AAC67 | MD 996(pAAC3-lacZY) | This study |
| AAC76 | MD 996(pAAC3-vcsJ2-lacZY) | This study |
| AAC78 | MD 996(pAAC3-vspD-lacZY) | This study |
| AAC87 | MD 996(pAAC3-vcsRTCNS2-lacZY) | This study |
| AAC89 | MD 996(pAAC3-vcsVU2-lacZY) | This study |
| AAC125 | MD 996(pAAC3-vopF-lacZY) | This study |
| AAC243 | MD992 integrated vcsRTCNS2-lacZY fusion | This study |
| AAC201 | MD992 integrated vspD-lacZY fusion | This study |
| AAC350 | MD992 integrated vcsVUQ2-lacZY fusion | This study |
| AAC355 | MD992 integrated vcsJ2-lacZY fusion | This study |
| AAC204 | MD992 integrated vopF-lacZY fusion | This study |
| AAC198 | MD992 integrated vttRB (A33_1675)-lacZY fusion | This study |
| AAC318 | MD992 integrated vttRA (A33_1664)-lacZY fusion | This study |
| AAC244 | MD992 integrated promoterless lacZY fusion | This study |
| AAC245 | AAC40 integrated vcsRTCNS2-lacZY fusion | This study |
| AAC259 | AAC40 integrated vspD-lacZY fusion | This study |
| AAC365 | AAC40 integrated vcsVUQ2-lacZY fusion | This study |
| AAC357 | AAC40 integrated vcsJ2-lacZY fusion | This study |
| AAC255 | AAC40 integrated vttRB (A33_1675)-lacZY fusion | This study |
| AAC321 | AAC40 integrated vttRA (A33_1664)-lacZY fusion | This study |
| AAC264 | AAC40 integrated promoterless lacZY | This study |
| AAC267 | AAC228 integrated vcsRTCNS2-lacZY fusion | This study |
| AAC282 | AAC228 integrated vspD-lacZY fusion | This study |
| AAC352 | AAC228 integrated vcsVUQ2-lacZY fusion | This study |
| AAC368 | AAC228 integrated vcsJ2-lacZY fusion | This study |
| AAC276 | AAC228 integrated vttRB (A33_1675)-lacZY fusion | This study |
| AAC324 | AAC228 integrated vttRA (A33_1664)-lacZY fusion | This study |
| AAC279 | AAC228 integrated promoterless lacZY | This study |
| AAC299 | AD10 integrated vcsRTCNS2-lacZY fusion | This study |
| AAC302 | AD10 integrated vspD-lacZY fusion | This study |
| AAC354 | AD10 integrated vcsVUQ2-lacZY fusion | This study |
| AAC358 | AD10 integrated vcsJ2-lacZY fusion | This study |
| AAC308 | AD10 integrated vttRB (A33_1675)-lacZY fusion | This study |
| AAC327 | AD10 integrated vttRA (A33_1664)-lacZY fusion | This study |
| AAC311 | AD10 integrated promoterless lacZY | This study |
| EK3 | MD1069 integrated vcsRTCNS2-lacZY fusion | This study |
| EK15 | MD1069 integrated vspD-lacZY fusion | This study |
| AAC486 | MD1069 integrated vcsVUQ2-lacZY fusion | This study |
| AAC490 | MD1069 integrated vcsJ2-lacZY fusion | This study |
| EK9 | MD1069 integrated vttRB (A33_1675)-lacZY fusion | This study |
| EK11 | MD1069 integrated vttRA (A33_1664)-lacZY fusion | This study |
| EK19 | MD1069 integrated promoterless lacZY | This study |
| E. coli | ||
| DH5αF′ | F′ endA1 hsdR17 supE44 thi-1 recA1 gyrA relA1 ΔU169(lacZYA-argF) (φ80 dlacΔM15) | Laboratory stock |
| SM10λpir | thi thr leu tonA lacY supE recA RP4-2-Tc::(λpir) Kanr | Laboratory stock |
| Plasmids | ||
| pAAC3 | Expression vector; Ampr | This study |
| pCVD442UMCS | Suicide vector with unique multiple cloning site, Ampr | Laboratory stock |
| pEH3 (EH077) | Allelic exchange suicide vector based on pCVD442; facilitates chromosomal integration of lacZY transcriptional fusions into V. cholerae lacZ; Ampr | This study |
| pCVD442UMCS-Δ vttRB | vttRB (A33_1675) deletion plasmid; Ampr | This study |
| pCVD442UMCS-Δ vttRA | vttRA (A33_1664) deletion plasmid; Ampr | This study |
| pCVD442UMCS-ΔtoxR | toxR deletion plasmid; Ampr | This study |
| pKB1 | phoA fusion vector based on pBSSK+; Ampr | This study |
| pKN1 | toxR-phoA fusion; Ampr | This study |
| pKN2 | vttRA-phoA fusion; Ampr | This study |
| pKN3 | vttRB-phoA fusion; Ampr | This study |
a
Ampr, ampicillin resistant; Strr, streptomycin resistant; Kanr, kanamycin resistant; R−, type II restriction endonuclease deletion; M+, methyltransferase positive; UMCS, unique multiple cloning site introduced into pCVD442 (MD1003).