Table 1

Strains and plasmids used in this study

Strain or plasmidRelevant characteristic(s)Source or reference
    E. coli
        LMG194Protein expression strainInvitrogen
        RosettaProtein expression strainNovagen
        S17-1recA endA thi hsdR RP4-2 Tc::Mu::Tn7 Smr70
        TOP10Cloning strainInvitrogen
    R. sphaeroides
        SF3flgT::aadAThis work
        SF4flgT::aadA ΔmotB1::KanThis work
        SF5flgT::aadA motBsup10 [MotB(S62P)]This work
        SF6flgT::aadA motBsup11 [MotB(F63S)]This work
        WS8Wild-type strain65
    S. cerevisiae AH109Yeast reporter strain for HIS3, ADE2, and lacZClontech
    pBAD/Myc-HisAExpression vector of His-tagged proteins; AprInvitrogen
    pBAD/Myc-HisA-flgTVector expressing FlgT-His6This work
    pGAD-flgTpGAD derivative expressing GAL4 AD-FlgTThis work
    pGAD-fliLppGADT7 derivative expressing GAL4 AD-FliLp56
    pGAD-motBppGADT7 derivative expressing GAL4 AD-MotBpThis work
    pGAD-motFppGADT7 derivative expressing GAL4 AD-MotFpThis work
    pGAD-motFpΔ77–98pGADT derivative expressing GAL4 AD MotFpΔ77–98This work
    pGAD-T7GAL4 activation domain; LEU2Clontech
    pGBD-flgTpGBD derivative expressing GAL4 BD-FlgTThis work
    pGBKT7GAL4 DNA binding domain; TRP1Clontech
    pJQ200mp18Mobilizable suicide vector; Gmr68
    pPIRLPlasmid that encodes tRNAs for rare codons; Cmr75
    pRK415Vector used for expression in R. sphaeroides; Tcr74
    pRK415_flgTpRK415 expressing FlgTThis work
    pRK_gfp-fliLpRK415 expressing the fusion GFP-FliL56
    pRK_gfp-motFpRK415 expressing the GFP-MotF fusion63
    pRKmotBsup2pRK415 expressing motBsup2 [MotB(A67E)]56
    pRKmotBsup4pRK415 expressing motBsup4 [MotB(F63L)]56
    pRKmotBsup5pRK415 expressing motBsup5 [MotB(A67D)]56
    pRKmotBsup6pRK415 expressing motBsup6 [MotB(A67T)]56
    pRKmotBsup8pRK415 expressing motBsup8 [MotB(A67G)]56
    pTZ19RCloning vector; AprPharmacia
    pTZ19R BampTZ19R derivative without BamHI siteLaboratory collection
    pTZ_FlgTflgT cloned into pTZ19R without BamHI siteThis work
    pWM5pUC derivative carrying the omega-Spcr cassette67