TABLE 1.

Strains and plasmids used in this study

Strain or plasmidRelevant characteristic(s)Source or reference
Strains
    DH5α recA endA1 gyrA96 hsd R17 thi-1 supE44 relA1 φ80dlacΖΔM15 Lab collection
    S17-1 thi pro hsdR recA chr::RP4-2 29
    GM2163F ara-14 leuB6 fhuA31 lacY1 tsx78 glnV44 galK2 galT22 mcrA dcm-6 hisG4 rfbD1 rpsL136 (Strr) dam13::Tn9 (Cmr) xylA5 mtl-1 thi-1 mcrB1 hsdR2 New England Biolabs
    PAO1Wild type P. aeruginosa strain ATCC 15692Lab collection
    PAO1ΔchpA PAO1 with Tcr insertion in chpA This study
Plasmids
    pBluescript II KS E. coli cloning vector with Ap selectionStratagene
    pOK12 E. coli cloning vector with Km selection 31
    pUC21 E. coli cloning vector with Ap selection 31
    pUCP19 E. coli-P. aeruginosa cloning vector 27
    pUCPSK P. aeruginosa-E. coli shuttle vector 32
    pMMB206 Pseudomonas controlled expression level vector with Cm selection 19
    pMOB3Counterselectable sacB/sacR genes from Bacillus subtilis 26
    pSM-TETSource of Tetr cartridge 18
    MTP26Cosmid containing P. aeruginosa PAO1 chromosomal DNA from bp 448166-472109 10
    pAL1pBluescript II KS with modified multiple cloning siteThis study
    pAL53,168-bp SacII/EcoRV fragment of chpA from MTP26 in pAL1This study
    pAL6458-bp H912i PCR product in pUC21This study
    pAL73,168-bp SacII/EcoRV fragment of chpA containing H1088 in pAL1This study
    pAL81,297-bp S1231i/H1410i PCR product in pUC21This study
    pAL9611-bp H1570i PCR product in pUC21This study
    pAL10872-bp BamHI/KpnI fragment of chpA containing H1717 in pUC21This study
    pAL111,396-bp EcoRV/KpnI fragment of chpA from MTP26 in pAL5This study
    pAL131,561-bp EcoRV/AatII fragment of chpA from MTP26 containing H54 in pOK12This study
    pAL14ChpA H54Q in pAL13This study
    pAL16ChpA H912Q in pAL6This study
    pAL17ChpA H1088Q in pAL7This study
    pAL18ChpA S1231A in pAL8This study
    pAL19ChpA H1410Q in pAL8This study
    pAL20ChpA H1570Q in pAL9This study
    pAL372,839-bp BamHI/ClaI fragment of chpA from MTP26 containing D2406N in pUC21This study
    pAL404,291-bp SacI/ClaI chpA fragment in pUCPSKThis study
    pAL41Full-length chpA under Plac control in pUCPSKThis study
    pAL42ChpA D2406N in pAL37This study
    pAL49Tcr cassette from pSM-TET (EcoRI/MluI) inserted into MfeI/AscI pAL11This study
    pAL525-kb NotI fragment from pMOB3 ligated into pAL49This study
    pAL577,972-bp NotI chpA fragment from MTP26 in pAL1This study
    pAL64pAL41 with an internal double FLAG tag inserted between the native NotI/NruI sites of chpA This study
    pAL773,862-bp ClaI/SacI chpA fragment from pAL41 in pUCP19This study
    pAL78Full-length chpA against Plac in pUCP19This study
    pAL798-kb ClaI/SacI fragment from MTP26 ligated into pMMB206 with PtaclacUV This study
    pAL823,503-bp StuI fragment of chpA cut from pAL57 and ligated into pAL14This study
    pAL83pAL79 with ChpA H54Q (mutation in HPt1 domain)This study
    pAL86pAL79 with ChpA H912Q (mutation in HPt2 domain)This study
    pAL87pAL79 with ChpA H1088Q (mutation in HPt3 domain)This study
    pAL88pAL79 with ChpA S1231A (mutation in SPt domain)This study
    pAL89pAL79 with ChpA H1410Q (mutation in HPt4 domain)This study
    pAL90pAL79 with ChpA H1570Q (mutation in HPt5 domain)This study
    pAL92pAL79 with ChpA D2406N (mutation in CheY domain)This study
    pAL79FLAGpAL79 with a double FLAG tag inserted into the NotI/KpnI sites of chpA This study
    pAL86FLAGpAL86 with a double FLAG tag inserted into the NotI/KpnI sites of chpA This study
    pAL87FLAGpAL87 with a double FLAG tag inserted into the NotI/KpnI sites of chpA This study
    pAL92FLAGpAL92 with a double FLAG tag inserted into the NotI/KpnI sites of chpA This study