TABLE 1.
Bacterial strains, plasmids, and oligonucleotide primers used in this study
| Strain, plasmid, or primer | Relevant genotype description or sequence (5′-3′) | Source or reference |
|---|---|---|
| V. cholerae strains | ||
| O395 | Wild type classical Ogawa | 30 |
| KKV598 | O395, ΔlacZ | 2 |
| KKV1113 | O395 ΔfliA ΔlacZ | 35 |
| KKV1247 | O395 ΔfliF ΔlacZ | This study |
| KKV1365 | O395 ΔfliA ΔflgM::Cm ΔlacZ | This study |
| KKV1384 | O395 ΔflgM ΔlacZ | This study |
| KKV1441 | O395 ΔflgM::Cm ΔlacZ | This study |
| KKV1461 | O395 ΔflgM ΔlacZ | This study |
| Plasmids | ||
| pKEK229 | R6K ori sacB mob Ampr | 2 |
| pKEK389 | ΔflgM in pKEK229 | This study |
| pKEK424 | ΔfliF in pKEK229 | 25 |
| pKEK427 | ΔflgM::Cm in pKEK229 | This study |
| pET15b | ColE1 ori His tag T7lac promoter Ampr | Novagen |
| pKEK460 | FLAG-flgM in pET15b | This study |
| pKEK462 | fliA in pET15b | This study |
| pKEK470 | p15A ori T7lac promoter, FLAG-flgM Cmr | This study |
| pBAD24 | ColE1 ori AmpraraBAD promoter | 10 |
| pKEK474 | FLAG-flgM in pBAD24 | This study |
| pRS551 | Transcriptional lacZ fusion vector Ampr Kanr | 38 |
| pKEK72 | flrB promoter-lacZ fusion in pRS551 | 21 |
| pKEK76 | flaC promoter-lacZ fusion in pRS551 | 20 |
| pKEK77 | flaD promoter-lacZ fusion in pRS551 | 20 |
| pKEK79 | flaB promoter-lacZ fusion in pRS551 | 20 |
| pKEK81 | flaE promoter-lacZ fusion in pRS551 | 20 |
| pKEK80 | flaA promoter-lacZ fusion in pRS551 | 20 |
| pKEK327 | fliE promoter-lacZ fusion in pRS551 | 35 |
| pKEK329 | flhA promoter-lacZ fusion in pRS551 | 35 |
| pKEK331 | flgK promoter-lacZ fusion in pRS551 | 35 |
| pKEK332 | flgB promoter-lacZ fusion in pRS551 | 35 |
| pKEK415 | flaG promoter-lacZ fusion in pRS551 | 35 |
| Primers | ||
| FLGMDEL1 | GCGGATCCGTGTGGGTGATGTGATTGAAC | |
| FLGMDEL2 | GCGAATTCGCTGGCATCAGAAGAACGCAC | |
| FLGMDEL3 | GCGAATTCGGCTCTTACGTGGTTGACCCT | |
| FLGMDEL4 | GCGAAGCTTCGGATTTGACTCACCAACTCA | |
| FLIANDEI | GGGGAACCTTCATATGAATAAAGCGC | |
| FLIAUBAMHI | GCGGATCCTTAGTCATTCCGAGTCCAAGAAC | |
| FLGMUBAMHI | GCGGATCCAGGTTAGAAGCCGCCCAGTTC | |
| FLAGFLGM | CGCGCCATGGACTACAAGGATGACGACGACAAGCATATGGCAGGTATTGATAACATACGC |