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. 2015 Oct 9;197(22):3499–3510. doi: 10.1128/JB.00419-15

TABLE 1.

Strains, plasmids, and oligonucleotides used in this study

Strain, plasmid, or oligonucleotide Relevant characteristics or sequence (5′ to 3′)a Source or reference
Strains
    E. coli
        EC100λpir supE44 ΔlacU169 (ϕ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1pirR6K) Epicentre
        SM10λpir thi-1 thr leu tonA lacY supE recA::RP4-2-Tc::mu KmrpirR6K) Lab collection
    V. cholerae
        JB58 O1 El Tor strain N16961 ΔlacZ Smr Lab collection
        XBV222 JB58 ΔleuO 23
        DT733 JB58 ΔtoxRS 23
        SS4 JB58 toxRΔppd This study
Plasmids
    pCM10 luxCDABE reporter plasmid, Kmr 51
    pJB906 pCM10 containing leuO promoter 23
    pTL61T lacZ transcriptional reporter plasmid, Cbr 52
    pXB266 pTL61T containing leuO promoter region with two ToxR binding sites
    pVA258 pTL61T containing leuO promoter region with one ToxR binding site This study
    pVA261 pTL61T containing leuO promoter region with no ToxR binding sites This study
    pXB233 pTL61T containing vexRAB promoter 53
    pXB228 pTL61T containing vexEF promoter 54
    pXB229 pTL61T containing vexGH promoter 54
    pXB230 pTL61T containing vexIJK promoter 54
    pXB231 pTL61T containing vexCD promoter 54
    pXB232 pTL61T containing vexLM promoter 54
    pBAD18 Arabinose-regulated expression plasmid, Cbr 55
    pBAD33 Arabinose-regulated expression plasmid, Cmr 55
    pXB298 pBAD18Km expressing leuO 23
    pVA94 pBAD18 expressing leuO This study
    pXB289 pBAD18 expressing toxRS 23
    pXB286 pBAD18 expressing toxRΔppdS 23
    pXB302 pBAD33 expressing toxRS This study
    pDT1391 pBAD33 expressing toxRΔppdS This study
    pWM91 Suicide plasmid vector used for allelic exchange 41
    pWM91::ΔtoxRppd pWM91 containing a fragment of toxR harboring a deletion of periplasmic domain This study
Oligonucleotides
    PleuO1-F CGCCCGGGAAATGCATTTTTATAGATTTTT
    PleuO2-F CGCCCGGGAATCGTATTGATTATTAAGGCT
    PleuO-R GGGGATCCGCGTCTTTTTTATCTAACATTTGCATGCCT
    toxRΔppd-F1 GGGAGCTCGGTCCTCAAAAGAGATAT
    toxRΔppd-F2 CTGCTCACTAACTAGGATCTTGCTAT
    toxRΔppd-R1 AACCCGGGCATGCCGCTCAGTCAGG
    toxRΔppd-R2 AGCAAGATCCTAGTTAGTGAGCAGTA
    5′BIO GCGGGAGTCGGCAGCG
    leuO-F-EMSA GCGGGAGTCGGCAGCGGTTAAAACATTTTTGACGTGAATATTAGTG
    leuO-R-EMSA GCGGGAGTCGGCAGCGCGTCACTAGCGATAAATATGCATAAATC
    ompU-F-EMSA GCGGGAGTCGGCAGCGCAATTAGATTGCGTGCATTT
    ompU-R-EMSA GCGGGAGTCGGCAGCGTTTTTTTACTCCCAAAGTTC
    vexR-F-EMSA GCGGGAGTCGGCAGCGTGCAAAACAGGGGGTATTAG
    vexR-R-EMSA GCGGGAGTCGGCAGCGGCCGTACACTATTTCAGACA
    leuO-qPCR-F GACCACTTCGCCACAAATCACCA
    leuO-qPCR-R CGTTGGATGGCGGAAAATGCG
    ompU-qPCR-F ACACCGTATAGGCTGTCATTG
    ompU-qPCR-R GTGCTGAAGCTCGCCTATCTC
    gyrA-qPCR-F CAATGCCGGTACACTGGTACG
    gyrA-qPCR-R AAGTACGGATCAGGGTCACG
a

Kmr, kanamycin resistant; Cbr, carbenicillin resistant; Cmr, chloramphenicol resistant.