TABLE 1.

Bacterial strains, plasmids, and primers used in this study

Strain, plasmid, or primerRelevant characteristics or sequence (5′-3′)aSource or reference
E. coli strains
    DH5αΔ(lacZYA-argF)U169 hsdR 17 relA 1 supE 44 endA 1 recA 1 thi-1 gyrA 96 φ80dlacZΔM15Life Technologies, Gaithersburg, MD
    DH5α λpir Δ(lacZYA-argF)U169 hsdR 17 relA 1 supE 44 endA 1 recA 1 thi-1 gyrA 96 φ80dlacZΔM15 λpir+ William W. Metcalf
    HB101F thi-1 hsdS 20(rB mB ) supE 44 recA 13 ara-14 proA 2 lacY 1 galK 2 rpsL 20 (Strr) xyl-5 mtl-1 40
P. aeruginosa strain
    PAO1Wild type 46
P. putida strains
    F1Wild type 18, 20
    PRS2000Wild type 37
    PRS4086PRS2000 cheA::mini-Tn5; Kanr 14
    F1 cheA F1 cheA::mini-Tn5; Kanr This study
    XLF004F1ΔPput_0623 (ΔmcpC)This study
    XLF028F1ΔPput_2527 (ΔcodA)This study
    XLF104F1ΔPput_0623 ΔPput_2527 (ΔmcpC ΔcodA)This study
Plasmids
    pAW19 sacB-containing cloning vector; Apr Kanr 53
    pRK2013ColE1 ori, RP4 mobilization function; Kanr 17
    pRK415Cloning vector, pRK290 derivative; Tetr 24
    pHJD215pRK415 carrying a 12-kb fragment that contains PRS2000 genes flhf, orfC, fliA, cheY, cheZ, and cheA with a mini-Tn5 insertion in cheA; Kanr Tetr 14
    pXLF004Gene Pput_0623 upstream and downstream 1-kb PCR products ligated and cloned into SpeI-SacI-digested pAW19; Apr Kanr This study
    pXLF204Gene Pput_0623 (mcpC) PCR product cloned into XbaI-EcoRI-digested pRK415; Tetr This study
    pXLFcodAGene Pput_2527 (codA) PCR product cloned into HindIII-EcoRI-digested pRK415; Tetr This study
    pXLFΔcodAGene Pput_2527 upstream and downstream 1-kb PCR products ligated and cloned into SpeI-digested pAW19; Apr Kanr This study
Primers
    0623 SpeI up-forGACTGTCACTAGTACCGCAGCAAACTCAATGTCThis study
    0623 up-rev (p)p-GCAACCGAAAATCAGAGCTTCThis study
    0623 dn-for (p)p-GATGCTCATCGACGCTCCCThis study
    0623 SacI dn-revGGACCTATGAGCTCCGATGTGCACGCCGAAGTGThis study
    0623 XbaI-forGTACATTCTAGACAGTTGCTACCTGCCTCGTGTGThis study
    0623 EcoRI-revGTACTCGAATTCACAGGCCATACGTCAGGTGCThis study
    CytoHindForATGCAAGCTTCGGTCGCCATGCTGGThis study
    CytoEcoRevATATGAATTCCCTGTGCACCTGAGGCGACThis study
    CytoSpeForGTGAACTAGTATTCGTCACCGGCATCAATCThis study
    UpCytoDELCACCTGAGGCGACAACAGCGGCTGTTACTCCTTGCCGTGGThis study
    DnCytoDELCGCTGTTGTCGCCTCAGGTGThis study
    CytoSpeRevGGTGACTAGTGCGTGAAGTCGGCCTACATCThis study
  • a Underlined bases indicate the positions of restriction sites. Apr, ampicillin resistant; Kanr, kanamycin resistant; Tetr, tetracycline resistant; (p) indicates that the primer carries a 5′ phosphate.