Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Reference or source |
|---|---|---|
| E. coli | ||
| E2348/69 | WT EPEC O127:H6 strain; Smr | 55 |
| ΔescN mutant | E2348/69 carrying an in-frame deletion of escN; Smr | 38 |
| ΔescV mutant | E2348/69 carrying an in-frame deletion of escV; Smr | 38 |
| Δler (JPEP24) mutant | E2348/69 carrying an in-frame deletion of ler; Kmr Smr | 8 |
| ΔescP mutant | E2348/69 carrying an in-frame deletion of escP; Kmr Smr | This study |
| BL21(DE3)/pLysS | Used for overproduction of proteins from pET-based plasmids; Cmr | Novagen |
| XL1-Blue | Used for cloning; Tcr | Stratagene |
| Salmonella | ||
| JR501 | For converting plasmid to Salmonella compatibility | 85 |
| SJW1103 | WT for motility and chemotaxis | 113 |
| S. cerevisiae AH109 | Yeast reporter strain; MATa trp1-901 leu2-3 ura3-52 his3-200 gal4Δ gal80Δ LYS2::GAL1UAS-GAL1TATA-HIS3 GAL2UAS-GAL2TATA-ADE2 URA3::MEL1UAS-MEL1TATA-lacZ | Clontech |
| Plasmids | ||
| pKD46 | Red recombinase system plasmid under control of the araB promoter | 15 |
| pKD4 | Template plasmid containing the kanamycin resistance cassette for the Red recombinase system | 15 |
| PCR-Blunt-II TOPO | Cloning vector | Invitrogen |
| pTrc99AFF4 | Modified pTrc99A expression vector | 80 |
| pTOPO-2HA | pCR2.1-TOPO derivative carrying C. rodentium espG coding region fused to two HA epitopes at the C terminus | 21 |
| pMAL-c2x | Expression vector for production of recombinant proteins with an N-terminal MBP tag | New England Biolabs |
| pET19b | T7 expression vector for production of recombinant proteins with an N-terminal His10 tag | Novagen |
| pET23b | T7 expression vector for production of recombinant proteins with a C-terminal His6 tag | Novagen |
| pGADT7 | Y2H vector containing GAL4 activation domain and LEU2 nutritional marker | Clontech |
| pGBKT7 | Y2H vector containing GAL4 DNA binding domain and TRP1 nutritional marker | Clontech |
| pGADT7-T | Encodes a fusion of the GAL4 activation domain with murine p53 | Clontech |
| pGBKT7-53 | Encodes a fusion of the GAL4 DNA binding domain with murine p53 | Clontech |
| pGBKT7-Lam | Encodes a fusion of the GAL4 DNA binding domain with human lamin C | Clontech |
| pJRo16 | pCR-Blunt II-TOPO carrying escP | This study |
| pJTo16 | pTrc99AFF4 carrying escP | This study |
| pMTBitirHcT | pTrc99AFF4 carrying tir and cesT with a His10 tag sequence at the 5′ end | Unpublished |
| pJHo16 | pTOPO-2HA carrying escP with its native RBS | This study |
| pJHΔ11o16 | pTOPO-2HA carrying Δ1-11escP with its native RBS | This study |
| pJHeH | pTOPO-2HA carrying espH with its native RBS | This study |
| pJHtir | pTOPO-2HA carrying tir with its native RBS | This study |
| pJHmap | pTOPO-2HA carrying map with its native RBS | This study |
| pJHnC | pTOPO-2HA carrying nleC with its native RBS | This study |
| pJHnD | pTOPO-2HA carrying nleD with its native RBS | This study |
| pJHnH | pTOPO-2HA carrying nleH with its native RBS | This study |
| pJHnI | pTOPO-2HA carrying nleI with its native RBS | This study |
| pJHeI | pTOPO-2HA carrying escI with its native RBS | This study |
| pJLo16 | pMAL-c2X carrying escP | This study |
| pJEeUC | pET19b carrying escUC | This study |
| pJEeUCC | pET19b carrying escUCC | This study |
| pJEeF | pET19b carrying escF | This study |
| pJEeI | pET19b carrying escI | This study |
| pMEcT | pET19b carrying cesT | Unpublished |
| pJEo16 | pET19b carrying escP | This study |
| pMEsL | pET19b carrying sepL | Unpublished |
| pJE23bo16 | pET23b carrying escP | This study |
| pOGADo16 | pGADT7 carrying escP | This study |
| pOGBKeUC | pGBKT7 carrying escUC | This study |
| pOGBKeUCC | pGBKT7 carrying escUCC | This study |
| pOGBKeF | pGBKT7 carrying escF | This study |
| pNGBKeI | pGBKT7 carrying escI | This study |
| pMGBKcT | pGBKT7 carrying cesT | Unpublished |
a
Sm, streptomycin; Km, kanamycin; Tc, tetracycline; Cm, chloramphenicol.