TABLE 1.

Strains, plasmids, and oligonucleotides used

Strain, plasmid, or oligonucleotideRelevant characteristic(s)aor sequenceDerivation or descriptionSource and/or reference
E. coli K-12
    DH5αF φ80dlacZΔM15 Δ(lacZYA-argF)U169 endA1 recA1 hsdR17(rK mK+) deoR thi-1 supE44 λ gyrA96 relA1Laboratory stock (61)
    SM10 (λpir)Rec RP4-2Tc::Mu λpirC60024, 65
    INVαF′F′ endA1 recA1 hsdR17(rK mK+) supE44 gyrA96 relA1 φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR thi-1 λInvitrogen
P. mirabilis
    BB2000Wild type, Rfr TcrSpontaneous from PRM113
    BB2021flgK2021::Tn5-Cm, Cmr Rfr Tcr, non-wild-type elongationMini-Tn5-CM insertion in BB2000This study
    BB2030flgL2030::Tn5-Cm, Cmr Rfr Tcr, non-wild type elongationMini-Tn5-CM insertion in BB2000This study
    BB2043fliF2234::Tn5-Cm, Cmr Rfr Tcr, non-wild type elongationMini-Tn5-CM insertion in BB2000This study
    BB2107fliM2107::Tn5-Cm, Cmr, Rfr Tcr, non-wild type elongationMini-Tn5-CM insertion in BB2000This study
    BB2109flgK2109::Tn5-Cm, Cmr, Rfr, Tcr, non-wild type elongationMini-Tn5-CM insertion in BB2000This study
    BB2122fliQ2122::Tn5-Cm, Cmr Rfr Tcr, non-wild type elongationMini-Tn5-CM insertion in BB2000This study
    BB2148fliG2148::Tn5-Cm, Cmr Rfr Tcr, non-wild type elongationMini-Tn5-CM insertion in BB2000This study
    BB2156flhA2156::Tn5-Cm, Cmr Rfr Tcr, non-wild type elongationMini-Tn5-CM insertion in BB2000This study
    BB2189flgK2189::Tn5-Cm, Cmr Rfr Tcr, non-wild-type elongationMini-Tn5-CM insertion in BB2000This study
    BB2193flgB2193::Tn5-Cm, Cmr Rfr Tcr, non-wild-type elongationMini-Tn5-CM insertion in BB2000This study
    BB2196flgK2196::Tn5-Cm, Cmr Rfr Tcr, non-wild-type elongationMini-Tn5-CM insertion in BB2000This study
    BB2199fliQ2199::Tn5-Cm, Cmr Rfr Tcr, non-wild-type elongationMini-Tn5-CM insertion in BB2000This study
    BB2201fliP2201::Tn5-Cm, Cmr Rfr Tcr, non-wild-type elongationMini-Tn5-CM insertion in BB2000This study
    BB2202flhA2202::Tn5-Cm, Cmr Rfr Tcr, non-wild-type elongationMini-Tn5-CM insertion in BB2000This study
    BB2203flhD2203::Tn5-Cm, Cmr Rfr Tcr, non-wild-type elongationMini-Tn5-CM insertion in BB2000This study
    BB2204fliL2204::Tn5-Cm, Cmr Rfr Tcr, non-wild-type elongationMini-Tn5-CM insertion in BB2000This study
Plasmids
    Vectors
        pACYC177Apr Kmrp15A ori low-copy-number plasmid vectorLaboratory stock
        pBluescriptIIAprpBluescriptII SK(+) and KS (−)Stratagene
        pBB402TcrpSB401 with deletion of EcoRI fragment containing Plux1This study
        pBB403Apr KmrpCR2.1 containing promoterless luxCDABE cassette amplified by PCR using 401FOR and 401REV with pBB402 templateThis study
        pBB404KmrpACYC177 with PstI fragment from pBB403 containing a promoterless luxCDABE cassette with an EcoRI cloning site 5′ to luxCThis study
        pBB413Kmr555-bp fragment bearing PzapA inserted upstream of luxCDABEThis study
        pBB415Kmr721-bp fragment bearing PhpmB inserted upstream of luxCDABEThis study
        pCR2.1Apr KmrPCR TOPO TA cloning vectorInvitrogen
        pRSI100Apr KmrpCR2.1 containing a 770-bp DNA containing 211 bp 5′ to the start of fliL and 76 bp 3′ to its stop codonInvitrogen
        pSB401TcrPromoterless luxCDABE cassette on pACYC18477
Oligodeoxyribonucleotides
    401FOR5′-CTGCAGGAATTCAGGCTTGGAGGATACGTATGACTAAAAAAATTTC-3′PCR primer for cloning the promoterless luxCDABE cassette from pBB402; contains PstI (underlined) and EcoRI (double underlined) sites at the 5′ endThis study
    401REV5′-CTGCAGGCGCTGATGTCCGGCGG-3′PCR primer for cloning the promoterless luxCDABE cassette from pBB402; contains a PstI site at the 5′ end (underlined)This study
    fliL-F5′-CTCTGCTCGTGGTGGTGTCG-3′PCR primer for cloning P. mirabilis fliLThis study
    fliL-R5′-GCGTCGTCACCTGATGTGTC-3′PCR primer for cloning P. mirabilis fliLThis study
    Tn5-I5′-AGATCTGATCAAGAGACAG-3′Primer for sequencing DNA flanking mini Tn5-CM insertions34, 51
    Tn5-O5′-ACTTGTGTATAAGAGTCAG-3′Primer for sequencing DNA flanking mini-Tn5-CM insertions34, 1
    zapApF5′-GGATCCAGATATTATTTTTATTAATAATAAGGATTAC-3′PCR primer for cloning the upstream regulatory region of zapA (PzapA) and containing a BamHI site at the 5′ end (underlined)This study
    zapApR5′-AATATCCTCCAGATATAAATTTAGGTTAAGTTTATT-3′PCR primer for cloning PzapAThis study
    hpmBpF5′-GGATCCTATGTAGTTTTTTTTTATCTATCATAAAC-3′PCR primer for cloning the upstream regulatory region of hpmB (PhpmB) and containing a BamHI site at the 5′ end (underlined)This study
    hpmBpR5′-TTTACCTCGAGATAAAGGTATTAATTCAA-3′PCR primer for cloning PhpmBThis study
This study
  • a Apr, Ampicillin resistance; Cmr, chloramphenicol resistance; Kmr, kanamycin resistance; Rfr, rifampin resistance; Tor, tetracycline resistance.