TABLE 1.
Strains and plasmids used in this studya
| Strain or plasmid | Description and/or genotype | Reference or source |
|---|---|---|
| E. tarda strains | ||
| PPD130/91 | Wild type; Kms Colr Amps; LD50 = 105.0 | 16 |
| ΔesaN strain | PPD130/91, in-frame deletion of esaN | 5 |
| ΔesaN/esaN strain | ΔesaN strain with pACYC-esaN | 5 |
| wt/GFP | PPD130/91 transformed with pFPV25.1 | This study |
| GFP-tagged ΔesaN strain | ΔesaN strain transformed with pFPV25.1 | This study |
| GFP-tagged ΔesaN/esaN strain | ΔesaN/esaN strain transformed with pFPV25.1 | This study |
| ΔfliC strain | PPD130/91, in-frame deletion of fliC | This study |
| ΔfliC/pACYC-fliC strain | ΔfliC strain with pACYC-fliC | This study |
| wt/pACYC184 | PPD130/91 with pACYC184 | This study |
| wt/pWSK29 | PPD130/91 with pWSK29 | This study |
| ΔfliC/pWSK29 strain | ΔfliC strain with pWSK29 | This study |
| ΔfliC/pWSK-fliC strain | ΔfliC strain with pWSK-fliC | This study |
| ΔflhB strain | PPD130/91, in-frame deletion of flhB | This study |
| ΔflhB ΔesaN strain | PPD130/91, in-frame deletion of flhB and esaN | This study |
| wt/pACYC-fliC2HA | PPD130/91 with pACYC-fliC2HA | This study |
| ΔflhB/pACYC-fliC2HA strain | ΔflhB strain with pACYC-fliC2HA | This study |
| ΔesaN/pACYC-fliC2HA strain | ΔesaN strain with pACYC-fliC2HA | This study |
| ΔflhB ΔesaN/pACYC-fliC2HA strain | ΔflhB ΔesaN strain with pACYC-fliC2HA | This study |
| E. coli strains | ||
| DH5α | α complementation | Stratagene |
| MC1061 λpir | (λpir) thi thr-1 leu6 proA2 his-4 argE2 lacY1 galK2 ara14 xyl5 supE44 λpir | 21 |
| S17-1 λpir | RK2 tra regulon, λpir, host for pir-dependent plasmid | 22 |
| Plasmids | ||
| pFPV25.1 | Derivative of pBR322 with gfpmut3A under the control of the constitutive promoter | 24 |
| pMD-18T | Ampr | TaKaRa |
| pACYC184 | Tetr Cmr | Amersham |
| pACYC-fliC | pACYC184 with wide-type fliC | This study |
| pACYC-fliC2HA | pACYC184 with C-terminal 2HA-tagged wild-type fliC | This study |
| pRE112 | pGP704 suicide plasmid, pir dependent; Cmr oriT oriV sacB | 19 |
| pRE112-fliCΔ1–416aa | pRE112 with fliC-flanking fragment, 1 to 416 amino acids deleted | This study |
| pRE112-flhBΔ4–430aa | pRE112 with flhB-flanking fragment, 4 to 430 amino acids deleted | This study |
| pWSK29 | Ampr | 23 |
| pWSK-fliC | pWSK29 with orf29/30 promoter and wild-type fliC | This study |
a
Km, kanamycin; Col, colistin; Amp, ampicillin; Cm, chloramphenicol; Tet, tetracycline; r, resistance; s, sensitivity.