TABLE 1.
Bacterial strains, plasmids, and primers
| Strain, plasmid, or primer | Genotype and/or description or sequence (5′–3′) | Reference |
|---|---|---|
| Strains | ||
| ECB480 | E22 (WT REPEC; serotype O103:H2) (Nalr) | 11 |
| VK550 | E22 ΔespZ (Nalr Kanr) | This work |
| VK581 | E22 ΔespZΩespZREPEC (Nalr Kanr); REPEC ΔespZ cis-complemented with REPEC espZ | This work |
| VK582 | E22ΔespZΩespZEPEC (Nalr Kanr); REPEC ΔespZ cis-complemented with EPEC espZ | This work |
| Plasmids | ||
| pMB196 | Vector backbone for cloning TEM-1 fusions | 25 |
| pMB201 | Vector control for translocation assays | 25 |
| pSR1 | Expresses the first 20 amino acids of EspZ fused to TEM-1 β-lactamase; used for translocation assays | This work |
| pSR2 | Plasmid for transient expression of REPEC espZ in eukaryotic cells; REPEC espZ allele cloned into pSR4 | This work |
| pSR3 | Plasmid for transient expression of EPEC espZ in eukaryotic cells; EPEC espZ allele cloned into pSR4 | This work |
| pSR4 | Lentiviral expression plasmid based on the pCIP-TTL-HA backbonea | This work |
| pGRG36 | Plasmid used to generate pJSW201 and pJSW101 | 24 |
| pJSW101 | Harbors EPEC espZ; used to generate VK582 | 18 |
| pJSW201 | Harbors REPEC espZ; used to generate VK581 | This work |
| Primers | ||
| Ni-EspZR | AGCGCAGTAACCCGGGGCTGCATCCATTAACTTTTCTCCA | |
| No-EspZ2 | TTTTGTCGACAGCGGGTCAATAACGCCTGC | |
| Ci-EspZ | TGGATGCAGCCCCGGGTTGGCGGCGGATTAGCGATGAAATATGC | |
| Co-EspZR2 | GTTCGTCGACGCCGTTAGCCAGTCACTATC | |
| KanF | CCCGGCATGCTAACTAGGAGGAATAAATG | |
| KanR | ATTACCCGTCCTGTCCTGAG | |
| RespZF | TGCAGGCTCTGAAGTAAGTAT | |
| RespZR | TTAGGCATATTTCATCGCTA | |
| SR1 | TCTAGAATGGAAGCAGCAAATTTAAGCCCTT | |
| SR2 | TCCGGAGGCATATTTCATCGCTAATCCGC | |
| SR3 | TCTAGAATGGATGCAGCAAATTTAAGCCCTT | |
| SR4 | TCCGGAGGCATATTTCATCGCTAATCCGC | |
| SR15 | TCATGAATTCCTGCAGGCTCTGAAGTAAG | |
| SR16 | TTCCGGTACCATTGATTGTGGCTGCCAGTG |
a
Kind gift from Felicia Goodrum, University of Arizona.