TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant feature(s) | Source or reference |
|---|---|---|
| Y. enterocolitica | ||
| 8081v | Wild type | 30 |
| JB580v | 8081v ΔyenR (R− M+) | 30 |
| GY357 | JB580v flhDC::pEP185.2 | This study |
| GY460 | JB580v flhDC | This study |
| JO1v | JB580v flgM::Kan | 29 |
| VK1 | JB580v fliA::Str | 29 |
| YCK10 | JB580v flgE::Cm | This study |
| YCK11 | JB580v flgF::Cm | This study |
| YMS12 | JB580v flhA::Cm | This study |
| YMS13 | JB580v flhB::Cm | This study |
| E. coli | ||
| LE392 | F−mcrA hsdR514 (R− M+) supE44 supF58 lacY1 galK2 galT22 metB1 trpRJS | 37 |
| DH5α | F′/endA1 hsdR17 (R− M+) supE44 thi-1 recA1 gyrA relA1 Δ(lacZYA-argF)U169, deoR [φ80d lacΔ(lacZ)M15] | 37 |
| MC4100 | F−araD139 Δ(lacZYA-argF)U169 rpsL150 relA1 flhD5301 deoC1 ptsF25 rbsR | 51 |
| MC1000 | F−araD139 galU galK Δ(lacZYA-argF)U169 rpsL150 | 47 |
| MC1000flhD::Kan | MC1000 but flhD::Kan | 47 |
| S17-1 λ(pir) | pro thi hsdR514 (R+ M−), ΔrecA, integrated RP4 2-Tc::Mu-Kn::Tn7 (Tpr Strr) | 52 |
| YK4131 | F−araD139 Δ(lacZYA-argF)U169 | 32 |
| Plsamids | ||
| pACYC177 | Medium-copy cloning vector, Knr | 37 |
| pACYC184 | Medium-copy cloning vector, Cmr Tetr | 37 |
| pBluescript KS+ | High-copy cloning vector | Stratagene |
| pEP185.2 | mob+, pir-dependent oriR6K, Cmr | 30 |
| pLAFR2 | Cosmid, Tetr | 11 |
| Litmus 28 | High-copy cloning vector | New England Biolabs |
| pRK2013 | mob+ tra+, self-transmissible plasmid, Kanr | 21 |
| pACYC4GM | pACYC184 containing a 4.0-kb flgDEF clone and a gentamicin resistance cassette | This study |
| pCKΩflgE | pBluescript KS+ with flgE::cat, nptI, and sacB | This study |
| pCKΩflgF | pBluescript KS+ with flgF::cat, nptI, and sacB | This study |
| pJB222 | pTM100 containing fliA, Tetr | 8 |
| pMG600 | pVLT33 with the S. liquefaciens flhDC locus cloned downstream of the ptac promoter as an EcoRI fragment, Kanr | 23 |
| pMSflh | pACYC177 with a 7-kb flhBAE clone | This study |
| pMSΩflhA | pBluescript KS+ with flhA::cat, nptI, and sacB | This study |
| pMSΩflhB | Litmus 28 with flhB::cat, nptI, and sacB | This study |
| pSWIM1 | pTM100 containing flgMN | 29 |
| PTM100 | mob+, derivative of pACYC184, Cmr Tetr | 39 |
| pVLT33 | mob+, low-copy vector containing an inducible ptac promoter, Kanr | 16 |
| pWKS130 | Low-copy cloning vector, ori pSC101, Kanr | 56 |
| pGY5-21 | pLAFR2 cosmid containing a ca. 10-kb insert of Y. enterocolitica chromosomal DNA containing the flhDC locus, Tetr | This study |
| pGY10 | pTM100 with a 4.3-kb EcoRI clone of flhDC originating from pGY5-22 | This study |
| pGY11 | ΔHpaI of pGY10 | This study |
| pGY12 | ΔSacII of pGY10 | This study |
| pGY13 | ΔBamHI of pGY15 | This study |
| pGY14 | ΔNcoI of pGY10 | This study |
| pGY15 | ΔEcoRV to BstZ17-I of pGY10 | This study |
| pGY16 | ΔEcoRI to PvuII of pGY15 | This study |
| pGY17 | ΔEcoRI to BstEII of pGY15 | This study |
| pGY18 | ΔEcoRI to SpeI of pGY15 | This study |
| pGY19 | pWKS130 with 1.4-kb BamHI flhDC fragment from pGY10 | This study |
| pGY20 | pVLT33 with 1.4-kb BamHI flhDC fragment from pGY10 placing flhDC under control of an inducible ptac promoter | This study |
| pGY21 | ca. 100-bp Sau3AI fragment from pGY10 cloned into the suicide vector pEP185.2 | This study |
| pGY22 | ca. 500-bp BamHI fragment encompassing the flhDC deletion from pGY11 cloned into the suicide vector pEP185.2 | This study |