Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant feature(s)Source or reference
Y. enterocolitica
 8081vWild type 30
 JB580v8081v ΔyenR (RM+) 30
 GY357JB580vflhDC::pEP185.2This study
 GY460JB580v flhDC This study
 JO1vJB580vflgM::Kan 29
 VK1JB580vfliA::Str 29
 YCK10JB580v flgE::CmThis study
 YCK11JB580v flgF::CmThis study
 YMS12JB580v flhA::CmThis study
 YMS13JB580v flhB::CmThis study
E. coli
 LE392F mcrA hsdR514 (R M+) supE44 supF58 lacY1 galK2 galT22 metB1 trpRJS 37
 DH5αF′/endA1 hsdR17 (RM+) supE44 thi-1 recA1 gyrA relA1Δ(lacZYA-argF)U169, deoR [φ80dlacΔ(lacZ)M15] 37
 MC4100F araD139Δ(lacZYA-argF)U169 rpsL150 relA1 flhD5301 deoC1 ptsF25 rbsR 51
 MC1000F araD139 galU galKΔ(lacZYA-argF)U169 rpsL150 47
 MC1000flhD::KanMC1000 butflhD::Kan 47
 S17-1 λ(pir) pro thi hsdR514 (R+M), ΔrecA, integrated RP4 2-Tc::Mu-Kn::Tn7 (TprStrr) 52
 YK4131F araD139Δ(lacZYA-argF)U169 32
Plsamids
 pACYC177Medium-copy cloning vector, Knr 37
 pACYC184Medium-copy cloning vector, CmrTetr 37
 pBluescript KS+High-copy cloning vectorStratagene
 pEP185.2 mob +, pir-dependentoriR6K, Cmr 30
 pLAFR2Cosmid, Tetr 11
 Litmus 28High-copy cloning vectorNew England Biolabs
 pRK2013 mob+ tra +, self-transmissible plasmid, Kanr 21
 pACYC4GMpACYC184 containing a 4.0-kb flgDEF clone and a gentamicin resistance cassetteThis study
 pCKΩflgE pBluescript KS+ withflgE::cat, nptI, andsacB This study
 pCKΩflgF pBluescript KS+ withflgF::cat, nptI, andsacB This study
 pJB222pTM100 containingfliA, Tetr 8
 pMG600pVLT33 with the S. liquefaciens flhDC locus cloned downstream of the ptac promoter as anEcoRI fragment, Kanr 23
 pMSflh pACYC177 with a 7-kb flhBAEcloneThis study
 pMSΩflhA pBluescript KS+ withflhA::cat, nptI, andsacB This study
 pMSΩflhB Litmus 28 with flhB::cat, nptI, andsacB This study
 pSWIM1pTM100 containingflgMN 29
 PTM100 mob +, derivative of pACYC184, Cmr Tetr 39
 pVLT33 mob +, low-copy vector containing an inducible ptac promoter, Kanr 16
 pWKS130Low-copy cloning vector, ori pSC101, Kanr 56
 pGY5-21pLAFR2 cosmid containing a ca. 10-kb insert of Y. enterocoliticachromosomal DNA containing the flhDC locus, Tetr This study
 pGY10pTM100 with a 4.3-kbEcoRI clone of flhDC originating from pGY5-22This study
 pGY11ΔHpaI of pGY10This study
 pGY12ΔSacII of pGY10This study
 pGY13ΔBamHI of pGY15This study
 pGY14ΔNcoI of pGY10This study
 pGY15ΔEcoRV to BstZ17-I of pGY10This study
 pGY16ΔEcoRI toPvuII of pGY15This study
 pGY17ΔEcoRI to BstEII of pGY15This study
 pGY18ΔEcoRI toSpeI of pGY15This study
 pGY19pWKS130 with 1.4-kb BamHI flhDC fragment from pGY10This study
 pGY20pVLT33 with 1.4-kb BamHIflhDC fragment from pGY10 placing flhDC under control of an inducible ptac promoterThis study
 pGY21ca. 100-bp Sau3AI fragment from pGY10 cloned into the suicide vector pEP185.2This study
 pGY22ca. 500-bp BamHI fragment encompassing the flhDCdeletion from pGY11 cloned into the suicide vector pEP185.2This study