TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant property(ies)a | Reference or origin |
|---|---|---|
| Strains | ||
| Yersinia pseudotuberculosis | ||
| 32777 | Wild-type strain | 28 |
| MYUH | Derived from strain 32777, ΔureHΩaphA-1a | This work |
| MYNT | Derived from strain 32777, ΔyntABCDEΩcat | This work |
| MYOU | Derived from strain 32777, ΔyntABCDEΩcat ΔureHΩaphA-1a | This work |
| Escherichia coli | ||
| DH5α | supE ΔlacU169 (φ80dlacZΔM15) thi deoR phoA hsdR recA endA gyrA relA | 16 |
| SY327λpir | Δ(lac pro) argE(Am) recA rif nalA λpir; host for pCVD442 and derivatives | 21 |
| SM10λpir | thi thr leu sup tonA lacY recA::RP4-2Tc::MuKm λpir; host for pCVD442 and derivatives | 32 |
| P4X | Hfr metB | 24 |
| HPX72 | Derived from strain P4X, nikA::MudI (Km lac) | 24 |
| Plasmids | ||
| pZero2-1 | Cloning vector, Km | Invitrogen |
| pUC18 | Cloning vector, Ap | 36 |
| pACYC184 | Plasmid vector, Cm Tc; source of the chloramphenicol acetyltransferase cat gene | 4 |
| pUC4K | Plasmid vector; Km; source of the aminoglycoside phosphotransferase aphA-1a gene | 34 |
| pCVD442 | Suicide vector containing the counterselectable marker sacB; Ap | 9 |
| pMS89 | pHC79Ω, ∼35-kb Sau3A DNA fragment from Y. pseudotuberculosis 32777 containing the urease locus and its flanking regions | 28 |
| pLW21 | pUC19Ω, 7.1-kb SmaI insert with the nikABCDER region from E. coli K-12 | 24 |
| pFS45 | pUC18Ω, ∼2.4-kb XhoI/Bst1107I insert with the 3′ end of yut and ureH from Y. pseudotuberculosis 32777 | This work |
| pFS50 | pACYC184Ω, ∼7.3-kb HindIII/PstI insert with the Y. pseudotuberculosis 32777 ure locus and its promoter region | This work |
| pFS78 | pUC18Ω, ∼5.5-kb PCR-generated fragment with primer set N1-N13 containing ynt genes from Y. pseudotuberculosis 32777 | This work |
| pFSU1 | pZero2-1Ω, ∼1.2-kb XbaI/EcoRI PCR-generated fragment with primer set N5-N6 encompassing the upstream region of ureH gene from Y. pseudotuberculosis 32777 | This work |
| pFSU2 | pFSU1Ω, ∼0.8-kb SacI/EcoRI PCR-generated fragment with primer set N7-N8 encompassing the downstream region of ureH gene from Y. pseudotuberculosis 32777 | This work |
| pFSU3 | pFSU2Ω, ∼1.3-kb EcoRI PCR-generated fragment with primer set K1-K2 encompassing aphA-1a from pUC4K | This work |
| pFSΔU | pCVD442Ω, ∼3.3-kb XbaI/SacI insert from pFSU3 | This work |
| pFSO1 | pUC18Ω, ∼0.5-kb HindIII/PstI PCR-generated fragment with primer set N1-N2 encompassing the upstream region of yntA gene from Y. pseudotuberculosis 32777 | This work |
| pFSO2 | pFSO1Ω, ∼0.5-kb BamHI/EcoRI PCR-generated fragment with primer set N3-N4 encompassing the downstream region of yntE gene from Y. pseudotuberculosis 32777 | This work |
| pFSO3 | pFSO2Ω, ∼1.4-kb PstI/BamHI PCR-generated fragment with primer set C3-C4 encompassing the cat gene from pACYC184 | This work |
| pFSΔO | pCVD442Ω, ∼2.4-kb SacI insert from pFSO3 | This work |
a
Ap, Cm, Km, and Tc, resistance to ampicillin, chloramphenicol, kanamycin, and tetracycline, respectively. Ω, in vitro insertion.