TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidGenotype or relevant characteristicsaReference or source
Strains
    P. aeruginosa
        PAO1Serotype O5; A+ B+ 21
        PAO1waaL waaL::Gmr A B derived from strain PAO1This work
        PAO1waaL + pUCP27-PAO1waaLPAO1waaL::Gmr complemented with pUCP27 having PAO1 waaLThis work
        PAO1waaL + pUCP27-PA14waaLPAO1waaL::Gmr cross-complemented with PA14 waaL in pUCP27This work
        PA14Serotype O10; A+ B+Fred Ausbel (Harvard Medical School)
        PA14waaL waaL::Gmr A B derived from strain PA14This work
        PA14waaL + pUCP27-PA14waaLPA14waaL::Gmr complemented with pUCP27 having PA14 waaLThis work
        PA14waaL + pUCP27-PAO1waaLPA14waaL::Gmr cross-complemented with PAO1 waaL in pUCP27This work
    E. coli
        JM109 recA1 supE44 endA1 hsdR17gyrA96 relA1 thi Δlac-proAB F′[traD36 proAB+lacIQlacZΔM15]
        SM10 thi-1 thr leu tonA lacY supE recA RP4-2-Tc::Mu Kmr 55
Plasmids
    pEX18APApr/CbrroriT+sacB+; gene replacement vector with multiple cloning site (MCS) from pUC18 27
    pPS856Gmr Apr; aacC1 gene (Gmr) from pUCP Gm ligated into the EcoRV site of pPS854; Gmr cassette is flanked by identical inverted MCS 27
    pUCP27pUC18-derived broad-host-range vector; Tcr 59
    pPAJL1PCR product of pa4999 upstream DNA (up to SalI site), cloned into pEX18AP using SmaI and SalI restriction sitesThis work
    pPAJL2PCR products of pa4999 downstream DNA (from SalI to 3′ end), cloned into pPA1 using SalI and PstI restriction sitesThis work
    pPAJL3PAO1 waaL knockout construct (insertion of Gmr cassette into pa4999)This work
    pPAJL4 pa4999 cloned into pUCP27 vector using SmaI and PstI restriction sitesThis work
    pPAJL5PCR product of PA14 waaL upstream DNA (up to SalI site), cloned into pEX18AP using SmaI and SalI restriction enzymes sitesThis work
    pPAJL6PCR products of PA14 waaL downstream DNA (from SalI to 3′ end), cloned into pPA1 using SalI and PstI restriction sitesThis work
    pPAJL7PA14 waaL knockout construct (insertion of Gmr cassette into PA14 waaL)This work
    pPAJL8PA14 waaL cloned into pUCP27 vector using SmaI and PstI restriction sitesThis work
  • a A superscript + or − sign after A or B designates the presence or absence of the particular O-polysaccharide.