TABLE 2.
Plasmids used in this study
| Plasmid | Characteristicsa | Reference or source |
|---|---|---|
| pEGFP-NleF | nleF from EPEC E2348/69 in pEGFP-C2, Kanr | 25 |
| pTrc99A | Bacterial expression vector with inducible lacI promoter, Ampr | Pharmacia Biotech |
| pTrc99A-NleF | nleF from EPEC E2348/69 in pTrc99A, Ampr | This study |
| pTrc99A-NleFΔLQCG | nleF from EPEC E2348/69 in pTrc99A with LQCG(186–189) motif deleted, Ampr | This study |
| pTrc99A-NleB1 | nleB1 from EPEC E2348/69 in pTrc99A, Ampr | 9 |
| pTrc99A-NleB1DXD | nleB1 from EPEC E2348/69 in pTrc99A with DXD(221–223) motif mutated to AAA, Ampr | 15 |
| p3XFlag-Myc-CMV-24 | Dually tagged N-terminal Met-3XFlag and C-terminal c-myc expression vector, Kanr | Sigma-Aldrich |
| pFlag-NleF | nleF from EPEC E2348/69 in p3XFlag-Myc-CMV-24, Kanr | This study |
| pFlag-NleB1 | nleB from EPEC E2348/69 in p3XFlag-Myc-CMV-24, Kanr | This study |
| pFlag-NleB1DXD | nleB from EPEC E2348/69 in p3XFlag-Myc-CMV-24 with DXD(221–223) motif mutated to AAA, Kanr | This study |
| pF_TRE3G_PGK puro | Lentiviral transduction vector with Tet-responsive element, Ampr | 34 |
| pF_TRE-Flag-NleF | nleF from EPEC E2348/69 with N-terminal 3XFlag tag in pF_TRE3G_PGK puro, Ampr | This study |
| pF_TRE-Flag-NleFΔLQCG | nleF from EPEC E2348/69 with N-terminal 3XFlag tag and LQCG(186–189) motif deleted in pF_TRE3G_PGK puro, Ampr | This study |
| pF_TRE-Flag-NleB1 | nleB1 from EPEC E2348/69 with N-terminal 3XFlag tag in pF_TRE3G_PGK puro, Ampr | This study |
| pF_TRE-Flag-NleB1DXD | nleB1 from EPEC E2348/69 with N-terminal 3XFlag tag and DXD(221–223) motif mutated to AAA in pF_TRE3G_PGK puro, Ampr | This study |
| pCMV-VSV-G | Mammalian expression vector encoding VSV-G glycoprotein, Ampr | John Silke |
| pCMVΔR8.2 | Mammalian expression vector encoding HIV-1 Gag/Pol, Tat, and Rev, Ampr | John Silke |
| pKD46 | Red recombinase helper plasmid, Ampr | 35 |
| pKD3 | Red recombinase helper plasmid, Cmr Ampr | 35 |
| pKD4 | Red recombinase helper plasmid, Kanr Ampr | 35 |
| pCR-Blunt II-TOPO | Cloning vector for blunt-end ligation, Kanr | Thermo Fisher Scientific |
| pCR-Blunt II-TOPO-NleBCR | nleB from C. rodentium ICC169 in pCR-Blunt II-TOPO with ribosomal s12 promoter and native NleB RBS, Kanr | This study |
| pCR-Blunt II-TOPO-NleFCR | nleF from C. rodentium ICC169 in pCR-Blunt II-TOPO with s12 promoter and RBS, Kanr | This study |
| pGRG36 | Tn7 transgene insertion vector, Ampr | 36 |
| pGRG36-NleBCR | nleB from C. rodentium ICC169 in pGRG36 with s12 promoter and native NleB RBS, Ampr | This study |
| pGRG36-NleFCR | nleF from C. rodentium ICC169 in pGRG36 with s12 promoter and RBS, Ampr | This study |
a
VSV, vesicular stomatitis virus; RBS, ribosome binding site.