TABLE 2

Bacterial strains and plasmids

Strain or plasmidDescriptionaReference or source
E. coli strains
    MG1655Wild-type K-1258
    MG1655ΔydeAK-12 MG1655ΔydeA::FRTThis work
    BL21F ompT hsdSB (rB mB) gal dcm59
    C41 (DE3)Derived from BL21(DE3): F ompT hsdSB (rB mB) gal dcm (DE3)60
    EG235C41 (DE3) ΔhipBA::kan, pMG25::gltX (optimized SD), pBAD33::His6::hipA (SD8 and start codon ATG)Laboratory collection
    JW2395AG1 [recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1] carrying pCA24N::gltX, GltX purification plasmid encoding N-terminally His6-tagged gltX, from ASKA collection55
Plasmids
    pBAD33p15 araC PBAD, Cmr61
    pNDM220Mini-R1 lacIq PA1/04/03, Ampr56
    pCP20pSC101 rep(Ts), Ampr Cmr47
    pBR322pMB1 rop, Ampr Tetr62
    pMG25pUC lacIq PA1/O4/O3, AmprLaboratory collection
    pEG25pMG25 derivative that has reduced leakiness of the IPTG-inducible PA1/O4/O3 promoterLaboratory collection
    pEG::gltXpMG25::gltX, optimized SDLaboratory collection
    pEG::His6hipApBAD33::His6hipA, HipA purification plasmid harboring N-terminally His6-tagged hipALaboratory collection
    pET-15bpBR322 lacI PT7, AmprNovagen
    pKG127pUC57::hipBSTO127Genscript
    pSVN1pBAD33::hipTO127, start codon GTGThis work
    pSVN42pEG25::hipTO127::His6, optimized SD, HipTO127 purification plasmid C-terminally His6-tagged hipTO127This work
    pSVN46pEG25::trpSHis6, optimized SD, TrpS purification plasmid harboring C-terminally His6-tagged trpSThis work
    pSVN49pEG25::trpSS197DHis6, optimized SD, TrpSS197D purification plasmid harboring C-terminally His6-tagged
trpSS197D
This work
    pSVN52pEG25::trpSS197AHis6, optimized SD, TrpSS197A purification plasmid harboring C-terminally His6-tagged
trpSS197A
This work
    pSVN60pUC57::His6-TEVhipB::hipS::hipTO127, optimized SDs for all genesGenscript
    pSVN61pUC57::hipB::hipS::hipTO127::His6, optimized SDs for all genes used for TA complex purification via
C-terminally His6-tagged hipTO127
Genscript
    pSVN37pEG25::trpS, optimized SDThis work
    pSVN44pBAD33::hipBSO127, optimized SD for hipBO127This work
    pSVN94pET-15b::His6-TEVhipB::hipS::hipTD233QO127, optimized SDs for all genesThis work
    pSVN103pNDM220::trpS, optimized SDThis work
    pSVN109pNDM220::hipSO127, optimized SDThis work
    pSVN110pNDM220::hipBSO127, optimized SDsThis work
    pSVN111pNDM220::hipBO127, optimized SDThis work
    pSVN113pUC57::hipBSTHiGenscript
    pSVN114pUC57::hipBSTTaGenscript
    pSVN116pNDM220::hipTO127, start codon GTGThis work
    pSVN122pNDM220::hipBHi, optimized SDThis work
    pSVN123pNDM220::hipSHi, optimized SDThis work
    pSVN124pNDM220::hipBSHi, optimized SD for hipBHi, overlapping hipBHi and hipSHiThis work
    pSVN126pNDM220::hipBTa, optimized SDThis work
    pSVN127pNDM220::hipSTa, optimized SDThis work
    pSVN128pNDM220::hipBSTa, optimized SD for hipBTa, overlapping hipBTa and hipSTaThis work
    pSVN129pBAD33::hipTTa, optimized SD, start codon GTGThis work
    pSVN135pBAD33::hipTHi, optimized SDThis work
    pSVN138pNDM220::hipBSTa, optimized SDsThis work
    pSVN139pNDM220::hipBSHi, optimized SDsThis work
    pAH1pNDM220::hipAA. Harms
    pAH2pNDM220::relEA. Harms
  • a SD, Shine-Dalgarno sequence.