TABLE 1.
Bacteria, plasmids, and phages used in this work
| Bacterial strains, plasmids, and phages | Description | Source or reference |
|---|---|---|
| Y. enterocolitica strains | ||
| 6471/76 (YeO3) | Serotype O:3, patient isolate, wild type | 37 |
| YeO3-Ail | Δail::Km-GenBlock, Kmr | This work |
| YeO3-Ail-R | Spontaneous rough derivative of YeO3-Ail, Kmr | This work |
| YeO3-Ail-OCR | Spontaneous OC mutant derivative of YeO3-Ail-R, Kmr | This work |
| YeO3-trs11 | Δ(wzx-wbcL)::Km-GenBlock, Kmr, derivative of YeO3 | 40 |
| YeO3-trs11-R | Spontaneous rough derivative of YeO3-trs11, Kmr | 40 |
| YeO3-OC | Δ(wzx-wbcQ), derivative of YeO3 | This work |
| YeO3-OCR | Spontaneous rough derivative of YeO3-OC | This work |
| YeO3-Ail-OC | Δail::Km-GenBlock, Kmr, derivative of YeO3-OC | This work |
| YeO3-R2 | Spontaneous rough derivative of YeO3 | 2 |
| YeO3-028 | ΔyadA::Km-GenBlock, Kmr, derivative of YeO3 | This work |
| YeO3-028-R | Spontaneous rough derivative of YeO3-O28, Kmr | This work |
| YeO3-028-OCR | Spontaneous OC mutant derivative of YeO3-028-R, Kmr | This work |
| YeO3-028-OC | Δ(wzx-wbcQ) derivative of YeO3-028, Kmr | This work |
| 6471/76-c (YeO3-c) | Virulence plasmid cured derivative of YeO3 | 37 |
| YeO3-R1 | Spontaneous rough derivative of YeO3-c | 2 |
| YeO3-c-OC | Δ(wzx-wbcQ) derivative of YeO3-c | This work |
| YeO3-c-OCR | Spontaneous rough derivative of YeO3-c-OC | This work |
| YeO3-c-trs8 | Δ(wzx-wbcL)::Km-GenBlock, Kmr, derivative of YeO3-c | 40 |
| YeO3-c-trs8R | Spontaneous rough derivative of YeO3-c-trs8, Kmr | 40 |
| YeO3-c-Ail | Δail::Km-GenBlock, Kmr, derivative of YeO3-c | This work |
| YeO3-c-Ail-OC | Spontaneous OC mutant derivative of YeO3-c-Ail, Kmr | This work |
| YeO3-c-Ail-R | Spontaneous rough derivative of YeO3-c-Ail, Kmr | This work |
| YeO3-c-Ail-OCR | Spontaneous OC mutant derivative of YeO3-c-Ail-R, Kmr | This work |
| E. coli strains | ||
| S17-1λpir | thi pro hsdR−hsdM+recA::RP4-2-Tc::Mu-Km::Tn7, Strr (λpir) | 13 |
| SY327λpir | Δ(lac pro) argE (Am) rif nalA recA56 (λpir) | 24 |
| HBI01/pRK2013 | Triparental conjugation helper strain, Kmr | 14 |
| C600 | thi thr leuB tonA lacY supE | 4 |
| Plasmids | ||
| pAil-8 | A pBR322 clone carrying an 8-kb insert of YeO3-c containing the ail gene; Ampr | This work |
| pAil-8.1 | PvuII deletion derivative of pAil-8; Ampr | This work |
| pAil-8.2 | EcoRV-NruI deletion derivative of pAil-8.1; Ampr | This work |
| pBR322 | Cloning vector; Ampr Tetr | 9 |
| pJM703.1 | Suicide vector, contains R6K origin of replication and RP4 Mob region; must be replicated in (λpir) hosts; Ampr | 24 |
| pRV1 | Suicide vector; derivative of pJM703.1; Clmr | 19 |
| pRV7 | A pBR322 clone carrying a 12.4-kb insert of YeO3-c covering the outer core gene cluster; Ampr | 40 |
| pRV37 | Intermediate to construct OC deletion suicide vector; pBR322 carries a HindIII fragment of pRV7 from which the OC gene cluster is deleted by PCR; Ampr | This work |
| pRV38 | 2-kb HindIII fragment of pRV37 cloned into pRV1 (O:3 OC gene cluster suicide vector); Clmr | This work |
| pRV42 | Suicide vector constructed to inactivate the ail gene; Km-GenBlock inserted between ail flanking fragments and cloned into pRV1; Kmr, Clmr | This work |
| pTM100 | Mobilizable vector, pACYC184-oriT of RK2; Clmr | 23 |
| pUC-4K | Origin of the Km-GenBlock cassette; Ampr, Kmr | Pharmacia |
| pYMS 4505 | yadA gene of YeO3 cloned into BamHI site of pTM100; Clmr | This work |
| pYMS 4506 | ΔyadA::Km-GenBlock derivative of pYMS4505; the internal 1.3-kb NsiI fragment of yadA gene replaced with Km-GenBlock; Clmr Kmr | This work |
| pYMS 4509 | ΔyadA::Km-GenBlock fragment cloned into EcoRI site of pJM703.1; Ampr Kmr | This work |
| pYMS 4515 | Suicide vector to inactivate the yadA gene; the cat gene of pTM100 cloned into the PstI site of pYMS4509; Clmr Kmr | This work |
| Bacteriophages | ||
| φYe03-12 | O-antigen-specific phage of Y. enterocolitica O:3 | 1 |
| φR1-37 | Outer core-specific phage of Y. enterocolitica O:3 | 40 |