TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| Strains | ||
| Y. enterocolitica | ||
| WA-314 | Clinical isolate of serotype O:8, carrying virulence plasmid pYVO8 | 18 |
| WA-c | Plasmidless derivative of WA-314 | 18 |
| E. coli | ||
| DH5α | endA1 supE44 hsdR17(rK− mK+) thi-1 recA1 gyrA96 relA1 Δ(lacZYA-argF)U169 (φ80 lacZΔM15) | 17 |
| Sm10λpir | thi-1 thr leu tonA lacY supE recA::RP4-2-TC::Mu-Kan (λpir), Kmr | 28 |
| ECOR-9 | Clinical isolate, EibA+ | 35, 45 |
| H. influenzae NTHi 58670 | Clinical isolate of a nontypeable H. influenzae strain, Hia+ | This study |
| M. catarrhalis O35E | Clinical isolate, UspA1+, UspA2+ | 2 |
| Plasmids | ||
| pUC-A-1 | pUC13 with 5-kb EcoRI-HindIII insert fragment of pYVO8 from WA-314 carrying the yadA gene | 43 |
| pUC-A-YadA-UspA1 | pUC-A1Δ334-422, carrying instead of the yadA linker and anchor region the homologous last 90 aa of UspA1 | This study |
| pUC-A-YadA-UspA1-2 | pUC-A1Δ362-422, carrying instead of the yadA loop and anchor region the homologous last 63 aa of UspA1 | This study |
| pUC-A-YadA-UspA1-3 | pUC-A1Δ369-422, carrying instead of the yadA anchor region the homologous last 56 aa of UspA1 | This study |
| pUC-A-YadA-EibA | pUC-A1Δ334-422, carrying instead of the yadA linker and anchor region the homologous last 87 aa of EibA | This study |
| pUC-A-YadA(D332L, H333E) | pUC-A1Δ334-422, carrying the reinserted yadA linker and anchor region | This study |
| pUC-A-YadA-Hia | pUC-A1Δ334-422, carrying instead of the yadA linker and anchor region the homologous last 90 aa of Hia | This study |
| pGP704 | Mobilizable suicide vector, R6K2 replicon, requires π protein in trans from the λpir-positive strain | 28 |
| pGPS-A-1 | pGP704 + 1.8-kb Spcr cassette in the EcoRI site (=pGPS) carrying the yadA gene as an EcoRI-SphI fragment from pUC-A-1 | 43 |
| pGPS-A-YadA-UspA1 | pGPS carrying the yadA-uspA1 gene as an EcoRI-SphI fragment from pUC-A-YadA-UspA1 | This study |
| pGPS-A-YadA-UspA1-2 | pGPS carrying the yadA-uspA1-2 gene as an EcoRI-SphI fragment from pUC-A-YadA-UspA1-2 | This study |
| pGPS-A-YadA-UspA1-3 | pGPS carrying the yadA-uspA1-3 gene as an EcoRI-SphI fragment from pUC-A-YadA-UspA1-3 | This study |
| pGPS-A-YadA-EibA | pGPS carrying the yadA-eibA gene as an EcoRI-SphI fragment from pUC-A-YadA-EibA | This study |
| pGPS-A-YadA(D332L, H333E) | pGPS carrying the yadA(D332L, H333E) gene as an EcoRI-SphI fragment from pUC-A-YadA(D332L, H333E) | This study |
| pGPS-A-YadA-Hia | pGPS carrying the yadA-hia gene as an EcoRI-SphI fragment from pUC-A-YadA-Hia | This study |
| pYVO8-A-0 | pYVO8, yadA mutant, Kmr cassette inserted in the PstI sites of yadA by allelic exchange | 43 |
| pYVO8-A-1 | pYVO8-A-0 with integrated pGPS-A-1, wild-type yadA | 43 |
| pYVO8-YadA-UspA1 | pYVO8-A-0 with integrated pGPS-A-YadA-UspA1 | This study |
| pYVO8-YadA-UspA1-2 | pYVO8-A-0 with integrated pGPS-A-YadA-UspA1-2 | This study |
| pYVO8-YadA-UspA1-3 | pYVO8-A-0 with integrated pGPS-A-YadA-UspA1-3 | This study |
| pYVO8-YadA-EibA | pYVO8-A-0 with integrated pGPS-A-YadA-EibA | This study |
| pYVO8-YadA(D332L, H333E) | pYVO8-A-0 with integrated pGPS-A-YadA(D332L, H333E) | This study |
| pYVO8-YadA-Hia | pYVO8-A-0 with integrated pGPS-A-YadA-Hia | This study |
a
Spcr, spectinomycin resistance; Kmr, kanamycin resistance.