Table 1

Strains and plasmids used in this study

Strain or plasmidRelevant characteristic(s)aReference or source
E. coli
    E2348/69WT EPEC O127:H6 strain; Smr55
    ΔescN mutantE2348/69 carrying an in-frame deletion of escN; Smr38
    ΔescV mutantE2348/69 carrying an in-frame deletion of escV; Smr38
    Δler (JPEP24) mutantE2348/69 carrying an in-frame deletion of ler; Kmr Smr8
    ΔescP mutantE2348/69 carrying an in-frame deletion of escP; Kmr SmrThis study
    BL21(DE3)/pLysSUsed for overproduction of proteins from pET-based plasmids; CmrNovagen
    XL1-BlueUsed for cloning; TcrStratagene
Salmonella
    JR501For converting plasmid to Salmonella compatibility85
    SJW1103WT for motility and chemotaxis113
S. cerevisiae AH109Yeast reporter strain; MATa trp1-901 leu2-3 ura3-52 his3-200 gal4Δ gal80Δ LYS2::GAL1UAS-GAL1TATA-HIS3 GAL2UAS-GAL2TATA-ADE2 URA3::MEL1UAS-MEL1TATA-lacZClontech
Plasmids
    pKD46Red recombinase system plasmid under control of the araB promoter15
    pKD4Template plasmid containing the kanamycin resistance cassette for the Red recombinase system15
    PCR-Blunt-II TOPOCloning vectorInvitrogen
    pTrc99AFF4Modified pTrc99A expression vector80
    pTOPO-2HApCR2.1-TOPO derivative carrying C. rodentium espG coding region fused to two HA epitopes at the C terminus21
    pMAL-c2xExpression vector for production of recombinant proteins with an N-terminal MBP tagNew England Biolabs
    pET19bT7 expression vector for production of recombinant proteins with an N-terminal His10 tagNovagen
    pET23bT7 expression vector for production of recombinant proteins with a C-terminal His6 tagNovagen
    pGADT7Y2H vector containing GAL4 activation domain and LEU2 nutritional markerClontech
    pGBKT7Y2H vector containing GAL4 DNA binding domain and TRP1 nutritional markerClontech
    pGADT7-TEncodes a fusion of the GAL4 activation domain with murine p53Clontech
    pGBKT7-53Encodes a fusion of the GAL4 DNA binding domain with murine p53Clontech
    pGBKT7-LamEncodes a fusion of the GAL4 DNA binding domain with human lamin CClontech
    pJRo16pCR-Blunt II-TOPO carrying escPThis study
    pJTo16pTrc99AFF4 carrying escPThis study
    pMTBitirHcTpTrc99AFF4 carrying tir and cesT with a His10 tag sequence at the 5′ endUnpublished
    pJHo16pTOPO-2HA carrying escP with its native RBSThis study
    pJHΔ11o16pTOPO-2HA carrying Δ1-11escP with its native RBSThis study
    pJHeHpTOPO-2HA carrying espH with its native RBSThis study
    pJHtirpTOPO-2HA carrying tir with its native RBSThis study
    pJHmappTOPO-2HA carrying map with its native RBSThis study
    pJHnCpTOPO-2HA carrying nleC with its native RBSThis study
    pJHnDpTOPO-2HA carrying nleD with its native RBSThis study
    pJHnHpTOPO-2HA carrying nleH with its native RBSThis study
    pJHnIpTOPO-2HA carrying nleI with its native RBSThis study
    pJHeIpTOPO-2HA carrying escI with its native RBSThis study
    pJLo16pMAL-c2X carrying escPThis study
    pJEeUCpET19b carrying escUCThis study
    pJEeUCCpET19b carrying escUCCThis study
    pJEeFpET19b carrying escFThis study
    pJEeIpET19b carrying escIThis study
    pMEcTpET19b carrying cesTUnpublished
    pJEo16pET19b carrying escPThis study
    pMEsLpET19b carrying sepLUnpublished
    pJE23bo16pET23b carrying escPThis study
    pOGADo16pGADT7 carrying escPThis study
    pOGBKeUCpGBKT7 carrying escUCThis study
    pOGBKeUCCpGBKT7 carrying escUCCThis study
    pOGBKeFpGBKT7 carrying escFThis study
    pNGBKeIpGBKT7 carrying escIThis study
    pMGBKcTpGBKT7 carrying cesTUnpublished
  • a Sm, streptomycin; Km, kanamycin; Tc, tetracycline; Cm, chloramphenicol.