Table 3.
Various bacterial strains and plasmids used in this study.
| Strain or Plasmid | Relevant Characteristic(s) | Component of | Source or Reference |
|---|---|---|---|
| A. hydrophila SSU | Diarrheal isolate of A. hydrophila | CDC* | |
| SSU-R | Rifampin resistant (Rifr) strain of A. hydrophila SSU | Laboratory Stock | |
| ΔascV | ascV mutant of A. hydrophila SSU-R; Rifr, Spr/Smr | T3SS | [61] |
| Δact | act mutant of A. hydrophila SSU-R; Rifr, Kmr | T2SS toxin | [60] |
| ΔaexU | aexU toxin mutant of A. hydrophila SSU-R; Rifr, Kmr | T3SS effector | [21] |
| ΔflhA | flhA mutant of A. hydrophila SSU-R; Rifr, Spr/Smr | Flagellar system | This study |
| ΔvasH | vasH mutant of A. hydrophila SSU-R; Rifr, Spr/Smr | T6SS | This study |
| ΔvasK | vasK mutant of A. hydrophila SSU-R; Rifr, Spr/Smr | T6SS | This study |
| Δact/ΔvasH | act and vasH double knockout mutant of A. hydrophila SSU-R; Rifr, Kmr, Spr/Smr | T2SS/T6SS | This study |
| Δact/ΔvasK | act and vasK double knockout mutant of A. hydrophila SSU-R; Rifr, Kmr,Spr/Smr | T2SS/T6SS | This study |
| ΔvasH/C | vasH mutant of A. hydrophila SSU-R complemented with pBR322-vasH; Rifr, Spr/Smr, Tcr | T6SS | This study |
| ΔvasK/C | vasK mutant of A. hydrophila SSU-R complemented with pBR322-vasK; Rifr,Spr/Smr, Tcr | T6SS | This study |
| WT/pBR322 | A. hydrophila SSU-R transformed with pBR322 vector alone; Rifr, Tcr, Apr | Laboratory Stock | |
|
| |||
| E.coli | |||
| DH5α | recA, gyrA | Life Technologies | |
| TOP10 | mcrA, hsdR, lacZ M15,endA1, recA1 | Invitrogen | |
| SM10 | λ pir | [60] | |
| HMS174 (DE3) | recA mutated K-12 background, that carries a chromosomal copy of the T7RNA polymerase under control of lacUV5 promoter. | Novagen | |
|
| |||
| Plasmids | |||
| pBR322 | Low copy plasmid used for complementation; Tcr, Apr | Pierce | |
| pET30a | pBR322-derived expression vector with T7 lac promotor, up- and down- stream His tags; Apr. | Novagen | |
| pDMS197 | Suicide vector; R6K ori, sacB, Tcr. | [62] | |
| pBI-EGFP | Eukaryotic expression vector regulated by a tetracycline element response (TER) and a minimal CMV promoter; Apr | Clontech | |
| pBluescript | Vector used for cloning proposes; Apr. | Stratagen | |
| pUC4K | Contains 1.2 Kb kanamycin resistance gene cassette; Kmr | Amersham | |
| pHPΩ45 | Contains a 2.0 Kb streptomycin/spectinomycin resistance gene cassette; Smr/Spr. | [63] | |
| pCR2.1 | Used for PCR cloning proposes; Apr, Kmr | Invitrogen | |
*
Centers for Diseases Control and Prevention