TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Genotype, description, or sequence | Reference or source |
|---|---|---|
| EPEC strains | ||
| E2348/69 | Wild-type 0127:H7 | |
| UMD874 | espF deletion mutant | 23 |
| E2348/69 cesF | cesF disruption mutant | 9 |
| EHEC strains | ||
| EDL933 | Wild-type O157:H7 Stx+ | |
| 85-170 | O157:H7 ΔStx | 37 |
| GH143 | cesF::Kan derivative of 85-170 | This study |
| GH145 | espF deletion derivative of 85-170 | This study |
| Plasmids | ||
| pUC 18 | Cloning vector | |
| pDEST17 | Cloning vector | Invitrogen |
| pKO3 | Counterselectable vector | 21 |
| pVK4 | Source of Kan cassette | 39 |
| pMP2 | Intermediate for pMP5 construction | This study |
| pMP3 | Intermediate for pMP4 construction | This study |
| pMP4 | Plasmid used to generate GH143 (EHEC ΔcesF) | This study |
| pMP5 | Plasmid used to generate GH145 (EHEC ΔespF) | This study |
| pIS4 | EPEC espF in pDEST17 to produce His-tag protein | This study |
| pVK166 | Full-length EHEC espF in pUC18 | This study |
| pSL1 | Full-length EHEC espF in pUC18 | This study |
| pVK167 | Full-length EHEC U-espF in pUC18 | This study |
| Primers | ||
| PF1 | GGCAGAATTCATGCTTAATGGAATTAGTAA | |
| PF3 | AATTCTGCAGTTACCCTTTCTTCGATTGCT | |
| PF6 | AAAAGCAGGCTTTATGCTTAATGGAATTAG | |
| PF7 | AGAAAGCTGGGTTTACCCTTTCTTCGATTGC | |
| PF10 | GGTAGAATTCATGATTAACAATGTTTCTTC | |
| PF11 | TATACTGCAGTCACGAGCGCTTAGATGTAT | |
| PF18 | NNNNCTGCAGTTAAATGCCATGCTCTGCAAGATG | |
| PF25 | GGGGCGGCCGCCTGGGTAATTGATCATGGTC | |
| PF26 | TCGACTAGTGATACCTACAAGCTGCCGCC | |
| PF27 | CATGAATTCGCCTATGAGCAATCGAAGAA | |
| PF28 | CACGTCGACCTGAATAACACATCCCCGTC | |
| PF29 | TGGCCTGCAGCATTAAAAATCGTCCTCGCA | |
| SF5 | ACAGGGATCCGGAGTTGGTTAGTCAGTTTG | |
| SF6 | ATGAGGATCCAGAGAGAAGGTGCTGGATTG | |
| 16S-F | ACTCAAATGAATTGACGGGGGC | |
| 16S-R | AGGCCCGGGAACGTATTCAC |