TABLE 1.
Bacterial strains, plasmids, and oligonucleotide primers used in this study
| V. cholerae strain, plasmid, or oligonucleotide | Relevant genotype or sequence (5′-3′)a | Source or reference or oligonucleotide used |
|---|---|---|
| Strains | ||
| KKV598 | 0395; ΔlacZ | 5 |
| KKV56 | 0395; ΔrpoN ΔlacZ | 21 |
| KKV98 | 0395; ΔflrC ΔlacZ | 21 |
| Plasmids | ||
| pRS551 | Transcriptional lacZ fusion vector; Ampr Kanr | 37 |
| pKEK75 | pRS551 with BglII and XbaI sites added | 21 |
| pKEK80 | −569 to +581 of flaAp in pRS551 | FLGLD1/FLAAP1 |
| pKEK147 | −54 to +581 of flaAp in pRS551 | FLAAPG2/FLAAP1 |
| pKEK158 | −285 to +347 of flaAp in pRS551 | FLAAPG4/FLAAPG3 |
| pKEK184 | −285 to +66 of flaAp in pRS551 | FLAAPG4/FLAAPG1 |
| pKEK445 | −54 to +206 of flaAp in pRS551 | FLAAPG2/FLAAPG5 |
| pKEK446 | −569 to +66 of flaAp in pRS551 | FLGLD1/FLAAPG1 |
| pKEK447 | −54 to +66 of flaAp in pRS551 | FLAAPG2/FLAAPG1 |
| pKEK594 | −54 to +137 of flaAp in pRS551 | FLAAPG2/FLAAPG7 |
| pKEK617 | −285 to +10 of flaAp in pRS551 | FLAAP10down, FLAAPG4like, FLAAP10up, FLAAPG5like |
| pKEK687 | −54 to +85 of flaAp in pRS551 | FLAAPG2/FLAAPG8 |
| pKEK691 | −54 to +95 of flaAp in pRS551 | FLAAPG2/FLAAP95 |
| pKEK692 | −54 to +110 of flaAp in pRS551 | FLAAPG2/FLAAPG9 |
| pKEK331 | −220 to +255 of flgKp in pRS551 | FLGKP1/FLGKP2 |
| pKEK580 | −220 to +27 of flgKp in pRS551 | FLGKP1/FLGKG2 |
| pKEK581 | −63 to +255 of flgKp in pRS551 | FLGKG1/FLGKP2 |
| pKEK657 | −63 to +207 of flgKp in pRS551 | FLGKG1/FLGKG4 |
| pKEK658 | −63 to +144 of flgKp in pRS551 | FLGKG1/FLGKG3 |
| pKEK667 | +11 to +144 flgKp fused to −220 to +10 flgKp in pRS551 | FLGKP220, FLGKP144, FLGKP10up, FLGKP10down |
| pKEK670 | −220 to +10 of flgKp in pRS551 | Same as pKEK667, digested with MfeI and BamHI to get rid of downstream region |
| pKEK695 | +24 to +110 flaAp fused upstream of −285 to +10 flaAp in pRS551 | FLAAP24, FLAAPG11, FLAAPG10, FLAAP10up |
| pKEK696 | +24 to +110 flaAp fused upstream of −220 to +10 flgKp in pRS551 | FLAAP24, FLAAK4, FLAAK3, FLGKP10up |
| pKEK697 | +11 to +144 flgKp fused upstream of −285 to +10 flaAp in pRS551 | FLGK10down, FLAAK2, FLAAK1, FLAAP10up |
| pKEK707 | +24 to +137 flaAp fused upstream of −285 to +10 flaAp in pRS551 | FLAAP24, FLAAPG13, FLAAPG12, FLAAP10up |
| pKEK714 | Same as pKEK707 with G+38→T in pRS551 | FLAAPG38T2/FLAAP10up |
| pKEK715 | Same as pKEK707 with G+78→T in pRS551 | FLAAP24, FLAAP88, FLAAP68, FLAAP10up |
| pKEK716 | Same as pKEK707 with G+38→T and G+78→T in pRS551 | FLAAP38T, FLAAP88, FLAAP68, FLAAP10up |
| Oligonucleotide | ||
| FLGLD1 | GCTCTAGAGGCTATCAGTTAGAGCGTAA | |
| FLAAP1 | GCAGATCTCGATTCATTCATCGCACCT | |
| FLAAPG1 | GCGGATCCAGCGGCAATCCATTTTGCCGC | |
| FLAAPG2 | GCTCTAGATATGAGCGAAGTGAGTTGAG | |
| FLAAPG3 | GCGGATCCGGTATTTACGTTAATGGTCATA | |
| FLAAPG4 | GCTCTAGAATTGGTGCGCGTTTGAGTACGC | |
| FLAAPG5 | GCGGATCCTACAACAACTCTGATGTACTTTA | |
| FLAAPG6 | GCGAATTCCACCGAAATCGGCAACTTTGAG | |
| FLAAPG7 | GCGGATCCCATCGCTTATTTTCAATGAGT | |
| FLAAPG8 | GCGGATCCAAAGTTGCCGATTTCGGTGAGC | |
| FLAAPG9 | GCGAATTCGGATCCGTCTATCTAATTCAATCAGC | |
| FLAAPG10 | TTAGATAGACATTGGTGCGCGTTTGAGTACG | |
| FLAAPG11 | GCGCACCAATGTCTATCTAATTCAATCAGC | |
| FLAAPG12 | ATAAGCGATGATTGGTGCGCGTTTGAGTACG | |
| FLAAPG13 | GCGCACCAATCATCGCTTATTTTCAATGAG | |
| FLAAP10down | GCGAATTCGAAAAGTTGTACAGGGTAACCA | |
| FLAAP10up | GCGGATCCACCAAGGCTCAATGTTGTATTAG | |
| FLAAPG4like | AGTTGTTGTACAATTGATTGGTGCGCGTTTGAGTAC | |
| FLAAPG5like | GCGCACCAATCAATTGTACAACAACTCTGATGTACT | |
| FLAAP24 | GCGAATTCGGGTAACCAAAGCGGCAAG | |
| FLAAP38T2 | GCGAATTCGGGTAACCAAAGCGTCAAGTCAGCG | |
| FLAAP68 | ACCGAAATCGTCAACTTTGAG | |
| FLAAP85 | GCGAATTCAAAGTTGCCGATTTCGGTGAGC | |
| FLAAP88 | CTCAAAGTTGACGATTTCGGT | |
| FLAAP95 | GCGAATTCGGATCCTCAGCCACTCAAAGTTGC | |
| FLAAP95GT | GCGAATTCGGATCCTCAGCCACTCAAAGTTGACGATTTC | |
| FLAAK1 | TAATACAGAGATTGGTGCGCGTTTGAGTACG | |
| FLAAK2 | GCGCACCAATCTCTGTATTAACATTGGAAAT | |
| FLAAK3 | TTAGATAGACCACCGTGCTGGATATGCGAC | |
| FLAAK4 | CAGCACGGTGGTCTATCTAATTCAATCAGC | |
| FLGKP1 | GCGAATTCCACCGTGCTGGATATGCGACC | |
| FLGKP2 | GCGGATCCTTGATCCCACGAGCGGCGAAC | |
| FLGKG1 | GCGAATTCTGCTATTTTTATTAGAGTAAATC | |
| FLGKG2 | GCGGATCCATTAATCAGTAAAATTCACTGAA | |
| FLGKG3 | GCGGATCCCTCTGTATTAACATTGGAAAT | |
| FLGKG4 | GCGGATCCGGTTTCACCGCCGTATTGGCG | |
| FLGKP10down | GCGAATTCGAATTTTACTGATTAATATCGG | |
| FLGKP10up | GCGGATCCACTGAATCTAAACAGTAGATAG | |
| FLGKP144 | CAGCACGGTGCAATTGCTCTGTATTAACATTGGAAATG | |
| FLGKP220 | TAATACAGAGCAATTGCACCGTGCTGGATATGCGACC |
a
Ampr, ampicillin resistance; Kanr, kanamycin resistance.