TABLE 2

Bacterial strains and plasmids used in this study

Strain or plasmidCharacteristics1Reference or source
Strains
    Escherichia coli BL21(DE3)F ompI hsdSB(rB mB) gal dam met69
        DH5αsupE44 lacU169(φ80lacZΔM15) hsdR17 (rK mK) recA1 endA1 gyrA96 thi-1 relA170
        HB101F Δ(gpt-proA)62 leuB6 supE44 ara-14 galK2 lacY1 Δ(mcrC-mrr) rpsL20 (Smr) xyl-5 mtl-1 recA13 thi-171
        JM109F′ traD36 proA+B+ lacIq Δ(lacZ)M15 Δ(lac-proAB) glnV44 e14 gyrA96 recA1 relA1 endA1 thi hsdR1772
        S17-1 λpirTpr Smr (recA thi pro hsdR) M+RP4: 2-Tc:Mu: Km Tn7 λpir73
    Ralstonia pseudosolanacearum Ps29Wild-type strain race 1, biovar 3, phylotype I74
    Pseudomonas putida KT2440Wild type15
        KT2440RRifampicin-resistant derivative of KT244075
        KT2440R-McpUKT2440R transposon mutant pp1228::mini-Tn5-Km; Rifr, Kmr76
    Pseudomonas aeruginosa PAO1Wild-type strain77
        PAO1 ΔpctAPAO1 derivative, pctA gene deletion mutantThis study
        PCTB1PAO1 derivative, pctB::Km; Km-23
        PCTC1PAO1 derivative, pctC::Km; Km-23
        PAO1 ΔtlpQPAO1 derivative, pa2654 gene deletion mutantThis study
        PCT2PAO1 derivative; ΔpctB-pctA-pa4308-pctC::km; Kmrb23
        PCTAQPAO1 derivative; ΔpctA ΔtlpQThis study
        PCT2QPAO1 derivative; ΔpctB-pctA-pa4308-pctC::Km, ΔtlpQ; KmrThis study
        PCT2QPPAO1 derivative; ΔpctB-pctA-pa4308-pctC::Km, ΔtlpQ ΔtlpP; KmrJ. Kato lab
        PCT2QARTPAO1 derivative; ΔpctB-pctA-pa4308-pctC, ΔtlpQ ΔtlpA (pa1646), ΔtlpR(pa2652) ΔtlpT(pa1930); KmrJ. Kato lab
    P. aeruginosa PA14Wild-type strain; human clinical isolate that elicits disease in plants, nematodes, insects, and mice78
        PA14-ACQPA14 derivative; ΔpctA ΔpctC ΔtlpQ Δpa4308This study
    P. aeruginosa isolate 227Isolated clinical strain from patients with urinary tract infections41
    P. aeruginosa isolate 233Isolated clinical strain from patients with urinary tract infections41
    P. aeruginosa isolate 287Isolated clinical strain from patients with urinary tract infections41
    P. aeruginosa isolate 401Isolated clinical strain from patients with urinary tract infections41
    P. aeruginosa isolate 428Isolated clinical strain from patients with urinary tract infections41
Plasmids
    pUCP18Escherichia-Pseudomonas shuttle vector; Apr79
    pPctApUCP18 with a PCR fragment containing pctA; Apr80
    pPctBpUCP18 with a PCR fragment containing pctB; Apr80
    pPctCpUCP18 with a PCR fragment containing pctC; AprThis study
    pTlpQpUCP18 with a PCR fragment containing tlpQ; Apr27
    pK18mobsacBPlasmid for allelic exchange; pK18 oriVE.c.lacZα mob sacB; Kmr81
    pK18mobsacB-pctApK18mobsacB containing a deletion of the pctA gene; KmrThis study
    pK18mobsacB-tlpQpK18mobsacB containing a deletion of the tlpQ gene; KmrThis study
    pK18mobsacB-pctABCpK18mobsacB containing a deletion of pctB, pctA, pa4308, pctC; KmrThis study
    pK18mobsacB-pctCpK18mobsacB containing a deletion of pctC and pa4308; KmrThis study
    pET28b(+)Protein expression plasmid; KmrNovagen
    pET28b-McpUpET28b derivative used to produce His-tagged McpU-LBD; Kmr20
    pET28b-TlpQpET28b derivative used to produce His-tagged TlpQ-LBD; KmrThis study
  • a Ap, ampicillin; Km, kanamycin; Rif, rifampin.

  • b The pa4308 gene (orf-1), which forms part of the pctABC operon, encodes a hypothetical protein that is not involved in chemotaxis (23).