TABLE 1

Strains and plasmids used in this study

Strain or plasmidCharacteristic(s)aReference or source
E. coli XL1-BluerecA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB laclqZΔM15 Tn10 (Tcr)]Stratagene
E. coli BL21(DE3)FompT gal dcm lon hsdSB (rB mB) with DE3, a λ prophage carrying the T7 RNA pol geneInvitrogen
A. tumefaciens strains
    A348C58 background carrying pTiA645
    AB320A348 with gaaPQM deletionThis study
    AB321A348 with udh deletionThis study
    AB335A348 with gaaR deletionThis study
    AB341AB900 with gaaP partial deletionThis study
    AB342AB901 with gaaP partial deletionThis study
    AB343AB900 with gaaPR148A mutationThis study
    AB344AB901 with gaaPR148A mutationThis study
    AB345AB900with gaaPR169A mutationThis study
    AB346AB901 with gaaPR169A mutationThis study
    AB347AB900 with gaaPR148Q mutationThis study
    AB348AB901with gaaPR148Q mutationThis study
    AB349AB900 with gaaPR169Q mutationThis study
    AB352AB901 with gaaPR169Q mutationThis study
    AB353AB900 with gaaPR148K mutationThis study
    AB354AB901 with gaaPR148K mutationThis study
    AB355AB900 with gaaPR169K mutationThis study
    AB356AB901 with gaaPR169K mutationThis study
    AB696A348 with gaaP deletionThis study
    AB900A348 but with RGS-6×His-tagged gaaPThis study
    AB901AB900 with gaaR deletionThis study
    AB902AB900 with hfq deletionThis study
Plasmids
    pET22bE. coli T7 expression vector; AprNovagen
    pBBR1MCS-4Broad-host-range plasmid cloning vector; Carbr46
    pBBR1MCS-5Broad-host-range plasmid cloning vector; Gmr46
    pK18mobsacBSuicide plasmid in Xanthomonas campestris pv. campestris; Kmr44
    pSW209ΩReporter plasmid carrying a PvirB::lacZ fusion; Specr48
    pJZ40pBBR1MCS-4 + gaaPQM; used for gaaPQM complementation; CarbrThis study
    pJZ41pBBR1MCS-4 + udh; used for udh complementation; CarbrThis study
    pJZ42pET22b + gaaR; used for gaaR overexpression in E. coli; AprThis study
    pJZ43pK18mobsacB derivative containing flanking sequences of gaaPQM; used for gaaPQM deletion; KmrThis study
    pJZ44pK18mobsacB derivative containing flanking sequences of gaaP; used for gaaP deletion; KmrThis study
    pJZ45pK18mobsacB derivative containing RGS-6×His-tagged gaaP and its flanking sequences; used for RGS-6×His-tagged gaaP recombination into chromosome; KmrThis study
    pJZ46pK18mobsacB derivative for deletion of 240 bp of the C terminus of gaaP; KmrThis study
    pJZ47pK18mobsacB derivative containing flanking sequences of udh; used for udh deletion; KmrThis study
    pJZ48pK18mobsacB derivative containing flanking sequences of gaaR; used for gaaR deletion; KmrThis study
    pJZ49pK18mobsacB derivative for R148A mutation of gaaP; KmrThis study
    pJZ50pK18mobsacB derivative for R148Q mutation of gaaP; KmrThis study
    pJZ51pK18mobsacB derivative for R148K mutation of gaaP; KmrThis study
    pJZ52pK18mobsacB derivative for R169A mutation of gaaP; KmrThis study
    pJZ53pK18mobsacB derivative for R169Q mutation of gaaP; KmrThis study
    pJZ54pK18mobsacB derivative for R169K mutation of gaaP; KmrThis study
    pJZ55pBBR1MCS-5 + hfq; used for hfq complementation; GmrThis study
  • a Tcr, tetracycline resistant; Apr, ampicillin resistant; Carbr, carbenicillin resistant; Gmr, gentamicin resistant; Kmr, kanamycin resistant; Specr, spectinomycin resistant.