TABLE 2.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant characteristic(s)Source or reference
Strains
    E. coli DH5αsupE44 ΔlacU16980lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1Clontech
    E. chrysanthemi
        Ech 3937Wild-type Saintpaulia (African violet) isolate27
        AC4150EC16 Nxr derivative, Chrysanthemum morifolium isolate12
        CUCPB5039AC4150 TTSS mutant5
        WPP42outD::kan; Kmr 3937 derivative; type II secretion mutantThis work
        WPP67hrpX::aadA; Spr/Smr 3937 derivativeThis work
        WPP92hrpY::kan; Kmr 3937 derivativeThis work
        WPP93hrpX::aadA hrpY::kan; Spr/Smr Kmr WPP67 derivativeThis work
        WPP90hrpS::cat; Cmr 3937 derivativeThis work
        WPP89hrpX::aadA hrpS::cat; Spr Cmr WPP67 derivativeThis work
        WPP96ΔhrpL(Δ1-185aa)::aadA; Spr/Smr 3937 derivativeThis work
        WPP98hrcJ::kan; Kmr 3937 derivativeThis work
        WPP100hrpA::cat; Cmr 3937 derivativeThis work
        WPP101bcsA::cat; Cmr 3937 derivativeThis work
        WPP102ΔhrpS(Δ46-212aa)::aadA; Spr/Smr 3937 derivativeThis work
        WPP119hrpX::aadA bcsA::cat; Spr/Smr Cmr WPP67 derivativeThis work
Plasmids
    pGEMT-EasyApr lacZ′ cloning vectorPromega
    pBluescript SK(+/−)Apr lacZ′ cloning vectorStratagene
    pQE31Apr His6-tagged expression vectorQiagen
    pHP45ΩSp/SmApr Spr/Smr broad-host-range cloning vector15
    pKD3Kmr template plasmid carrying kan cassette13
    pKD4Cmr template plasmid carrying cat cassette13
    pJN105Gmr, arabinose inducible expression vector34
    pCPP50Apr pINIII113-A2-based expression vector8
    p519ngfpKmr broad-host-range vector31
    pRK415Tcr broad-host-range low-copy vector24
    pThrpYLΩSpApr Spr/Smr pGEMT-Easy carrying 4.8-kb hrpY-hrpL region containing hrpX::ΩSp insertion at StuI siteThis work
    pThrpYLKmApr Kmr pGEMT-Easy carrying 4.4-kb hrpY-hrpL region containing hrpY::Km insertion at blunt-ended BglII siteThis work
    pThrcJY::catApr Kmr pGEMT-Easy carrying 4.4-kb hrcJ-hrpY region containing hrpS::Cm insertion at PstI siteThis work
    pThrpYJΩSpApr 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 ΩSpThis work
    pThrcJYKmApr Kmr pGEMT-Easy carrying 5-kb hrcJ-hrpY region containing hrcJ::Km insertion at SmaI siteThis work
    pBShrc/pJSCmApr Cmr pBluescript SK(+/−) carrying 3-kb hrcJ-hrpS containing hrpA::Cm insertion at blunt-ended SphI siteThis work
    p519nGFPbcsAKmr Cmr p519ngfp with bcsA::cat inserted into EcoRI site; cat is inserted into the BamHI site of bcsAThis work
    pYAΔSΩSpApr Spr 2.0-kb hrpS-deleted hrpY-hrpA region on pGEMT-Easy; hrpS was removed using EcoRV and StuI and replaced with an ΩSp cassetteThis work
    p105XYGmr 2.5-kb hrpXY with native promoter cloned into BamHI and XbaI sites of pJN105This work
    pQhrpYApr 0.7-kb hrpY in BamHI and HindIII sites of pQE31This work
    p105SGmr 1.5-kb hrpS with native promoter cloned into BamHI and XbaI sites of pJN105This work
    p50hrpLApr 0.55-kb hrpL in SpeI and EcoRI sites of pCPP50This work
    p50hrcJApr 0.97-kb hrcJ in SpeI and EcoRI sites of pCPP50This work
    p415hrpATcr 0.3-kb hrpA cloned into XbaI and EcoRI sites of pRK415This work
    p50bcsAApr 3-kb bcsA with native promoter cloned into pCPP50This work