TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | F− λ− recA1 hsdR17 endA1 supE44 thi-1 relA1 gyrA96 Δ(argF-lacZYA)U169 ϕ80dlacZΔM15 (recipient in cloning experiments) | 32 |
| S17-1 | recA hsdR thi pro ara RP-4 2-Tc::Mu-Km::Tn7 (Tpr Smr) (recipient in conjugational transfer of pMMB206) | 33 |
| β3914 | β2163 gyrA462 zei-298::Tn10 (Kmr Emr Tcr) (recipient in conjugational transfer of pSW7848) | 34 |
| BL21(DE3) | Host for overexpression | Stratagene |
| V. alginolyticus strains | ||
| VIO5 | Rifr Pof+ Laf− | 35 |
| LPN1 | VIO5 ΔflhF | 7 |
| KK148 | VIO5 flhG | 6 |
| LPN2 | VIO5 ΔflhFG | 7 |
| NMB303 | VIO5 ΔhubP | This study |
| NMB304 | LPN1 ΔhubP | This study |
| NMB305 | KK148 ΔhubP | This study |
| NMB308 | VIO5 hubP-egfp | This study |
| Plasmids | ||
| pGEM-T Easy | Cloning vector, Ampr | Promega |
| pTSK117-1 | pGEM-T Easy-hubP* | This study |
| pTSK117 | pGEM-T Easy-hubP*-egfp | This study |
| pSW7848 | Suicide vector (oriVR6Kγ oriTRP4 araC-PBAD-ccdB), Cmr | 36 |
| pTSK118 | pSW7848-hubP*-egfp | This study |
| pSJ2 | pSW7848-ΔhubP | This study |
| pBAD33 | Cmr PBAD | 31 |
| pTY200 | pBAD33-egfp | 37 |
| pTSJ3 | pBAD33-hubP | This study |
| pTSJ4 | pBAD33-hubP-His6 | This study |
| pAK520 | pBAD33-flhG | 6 |
| pAK325 | pBAD33-flhF-egfp | 7 |
| pAK541 | pBAD33-flhG-egfp | 7 |
| pTY60 | Kmr PBAD | 38 |
| pTY60-flhF-gfp | pAK325, Cms, Kmr | Akiko Kusumoto |
| pMMB206 | Cmr Ptac Plac UV5 | 39 |
| pNT16 | pMMB206-flhG | This study |
| pTrcHisB | Ampr Ptrc | Invitrogen |
| pTrc-flhG | pTrcHisB-His6-tev-flhG | 17 |
a
Tpr, trimethoprim resistant; Smr, streptomycin resistant; Kmr, kanamycin resistant; Emr, erythromycin resistant; Tcr, tetracycline resistant; Rifr, rifampin resistant; Ampr, ampicillin resistant; Cmr, chloramphenicol resistant; Cms, chloramphenicol sensitive; Pof+, normal polar flagellar formation; Laf−, defective in lateral flagellar formation; Ptac, tac promoter; Plac, lac promoter; PBAD, arabinose promoter; Ptrc, trc promoter; hubP*-egfp, ∼1.7-kb DNA fragment containing the hubP 3′ coding region and its downstream sequence, with an in-frame insertion of the egfp sequence at the 3′ end of hubP.