TABLE 2.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| E. coli DH5α | supE44 ΔlacU169 (Δ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Clontech |
| E. chrysanthemi | ||
| Ech 3937 | Wild-type Saintpaulia (African violet) isolate | 27 |
| AC4150 | EC16 Nxr derivative, Chrysanthemum morifolium isolate | 12 |
| CUCPB5039 | AC4150 TTSS mutant | 5 |
| WPP42 | outD::kan; Kmr 3937 derivative; type II secretion mutant | This work |
| WPP67 | hrpX::aadA; Spr/Smr 3937 derivative | This work |
| WPP92 | hrpY::kan; Kmr 3937 derivative | This work |
| WPP93 | hrpX::aadA hrpY::kan; Spr/Smr Kmr WPP67 derivative | This work |
| WPP90 | hrpS::cat; Cmr 3937 derivative | This work |
| WPP89 | hrpX::aadA hrpS::cat; Spr Cmr WPP67 derivative | This work |
| WPP96 | ΔhrpL(Δ1-185aa)::aadA; Spr/Smr 3937 derivative | This work |
| WPP98 | hrcJ::kan; Kmr 3937 derivative | This work |
| WPP100 | hrpA::cat; Cmr 3937 derivative | This work |
| WPP101 | bcsA::cat; Cmr 3937 derivative | This work |
| WPP102 | ΔhrpS(Δ46-212aa)::aadA; Spr/Smr 3937 derivative | This work |
| WPP119 | hrpX::aadA bcsA::cat; Spr/Smr Cmr WPP67 derivative | This work |
| Plasmids | ||
| pGEMT-Easy | AprlacZ′ cloning vector | Promega |
| pBluescript SK(+/−) | AprlacZ′ cloning vector | Stratagene |
| pQE31 | Apr His6-tagged expression vector | Qiagen |
| pHP45ΩSp/Sm | Apr Spr/Smr broad-host-range cloning vector | 15 |
| pKD3 | Kmr template plasmid carrying kan cassette | 13 |
| pKD4 | Cmr template plasmid carrying cat cassette | 13 |
| pJN105 | Gmr, arabinose inducible expression vector | 34 |
| pCPP50 | Apr pINIII113-A2-based expression vector | 8 |
| p519ngfp | Kmr broad-host-range vector | 31 |
| pRK415 | Tcr broad-host-range low-copy vector | 24 |
| pThrpYLΩSp | Apr Spr/Smr pGEMT-Easy carrying 4.8-kb hrpY-hrpL region containing hrpX::ΩSp insertion at StuI site | This work |
| pThrpYLKm | Apr Kmr pGEMT-Easy carrying 4.4-kb hrpY-hrpL region containing hrpY::Km insertion at blunt-ended BglII site | This work |
| pThrcJY::cat | Apr Kmr pGEMT-Easy carrying 4.4-kb hrcJ-hrpY region containing hrpS::Cm insertion at PstI site | This work |
| pThrpYJΩSp | Apr Spr 5.2-kb hrpL-deleted hrpY-hrpJ region on pGEMT-Easy; complete hrpL coding region was removed with SmaI and HindIII, blunt ended, and ligated to ΩSp | This work |
| pThrcJYKm | Apr Kmr pGEMT-Easy carrying 5-kb hrcJ-hrpY region containing hrcJ::Km insertion at SmaI site | This work |
| pBShrc/pJSCm | Apr Cmr pBluescript SK(+/−) carrying 3-kb hrcJ-hrpS containing hrpA::Cm insertion at blunt-ended SphI site | This work |
| p519nGFPbcsA | Kmr Cmr p519ngfp with bcsA::cat inserted into EcoRI site; cat is inserted into the BamHI site of bcsA | This work |
| pYAΔSΩSp | Apr Spr 2.0-kb hrpS-deleted hrpY-hrpA region on pGEMT-Easy; hrpS was removed using EcoRV and StuI and replaced with an ΩSp cassette | This work |
| p105XY | Gmr 2.5-kb hrpXY with native promoter cloned into BamHI and XbaI sites of pJN105 | This work |
| pQhrpY | Apr 0.7-kb hrpY in BamHI and HindIII sites of pQE31 | This work |
| p105S | Gmr 1.5-kb hrpS with native promoter cloned into BamHI and XbaI sites of pJN105 | This work |
| p50hrpL | Apr 0.55-kb hrpL in SpeI and EcoRI sites of pCPP50 | This work |
| p50hrcJ | Apr 0.97-kb hrcJ in SpeI and EcoRI sites of pCPP50 | This work |
| p415hrpA | Tcr 0.3-kb hrpA cloned into XbaI and EcoRI sites of pRK415 | This work |
| p50bcsA | Apr 3-kb bcsA with native promoter cloned into pCPP50 | This work |