Table 1. Bacterial strains and plasmids used in this study.
| Strains or plasmids | Relevant properties | Reference or source | |
| E. coli | |||
| SM10lacIq | thi thr leu tonA lacY supE recA::RP4-2-Tc::Mu Km lacIq | T. Hoang | |
| SM10 (λpir) | thi thr leu tonA lacY supE recA::RP4-2-Tc::Mu Km λpir | [15] | |
| DL291 | F- araD139 Δ(argF-lac)U169 rpsL150 deoC1 relA1 rbsR ptsF25 flbB5301 glpR2 gyrA Δ(glpT-glpA)593 recA1 | [18] | |
| DL291-Tn7 b | Gmr; DL291 derivative containing mini-Tn7T-fabA′-′lacZY gene fusion | This study | |
| P. aeruginosa | |||
| PAO1 | Prototroph | [36] | |
| PAO434 | PAO1 with mini-Tn7T-lacZ | This study | |
| PAO435 | PAO1 with mini-Tn7T-pfabA′-lacZ a resulted from integration of pPS1460 | This study | |
| PAO459 | PAO1 with mini-Tn7T-pfabA′-lacZ resulted from integration of pPS1491 | This study | |
| PAO460 | PAO1 with mini-Tn7T-pfabAΔ30′-lacZ | This study | |
| PAO483 | PAO434 with ΔPA4890::FRT a | This study | |
| PAO484 | PAO435 with ΔPA4890::FRT | This study | |
| PAO491 | PAO434 with ΔPA1539::FRT | This study | |
| PAO492 | PAO435 with ΔPA1539::FRT | This study | |
| PAO495 | PAO460 with ΔPA4890::FRT | This study | |
| PAO497 | PAO460 with ΔPA1539::FRT | This study | |
| PAO517 | PAO435 with ΔPA4890::FRT ΔPA1539::FRT | This study | |
| PAO194 | PAO1 with temperature-sensitive fabA(Ts) | This study | |
| PAO1010 | PAO435 with Δanr::FRT | This study | |
| PAO1151 | Gmra; PAO1 with mini-Tn7T-pPA1612′-lacZ | This study | |
| Plasmid | |||
| pPS752 | Apr; fabA+ fabB + (4.8 kb chromosomal BamHI-EcoRI fragment cloned between the same sites of pUC18) | [2] | |
| pPS790 | Apr; fabA+ fabB + (2,039 bp PCR amplified BamHI-KpnI fragment from pPS752 cloned between the same sites of pUC19) | [2] | |
| pPS1450 | Apr, Gmr; mini-Tn7 delivery vector | [37] | |
| pPS1453 | Apr, Gmr; mini-Tn7 delivery vector with a promoter-less lacZ gene | [37] | |
| pPS1460 | Apr, Gmr; ligation of ∼300 bp SalI-SmaI fragment from pPS790 to XhoI-NruI fragment of pPS1453 | This study | |
| pPS1482 | Apr; pCR2.1 with PCR-amplified PA1611-fabA intergenic region from PAO1 | This study | |
| pPS1488b | Apr; pCR-Blunt II-TOPO with PCR-amplified PA1611-fabA intergenic containing a 30 bp deletion | This study | |
| pPS1491 | Apr, Gmr; pUC18-mini-Tn7T-pfabA′-lacZ delivery vector obtained by ligation of ∼230 bp KpnI-SmaI fragment of pPS1482 into the same sites of pPS1453 | This study | |
| pPS1492 | Apr, Gmr; pUC18-mini-Tn7T-pfabAΔ30::lacZ delivery vector obtained by ligation of ∼230 bp KpnI-SmaI fragment of pPS1488 into the same sites of pPS1453 | This study | |
| pDONR221 | Kmr; Gateway donor vector | Invitrogen | |
| pEX18Ap | Apr; gene replacement vector with MCS from pUC18 | [13] | |
| pEX18ApGW | Apr; Gateway destination vector | [14] | |
| pPS1669 | Apr, Gmr; pUC18-mini-Tn7T-Gm-lacZ-GW | [14] | |
| pFLP2 | Apr; source for Flp recombinase | [13] | |
| pPS856 | Apr, Gmr; Gm-FRT cassette | [13] | |
| pBT20 | Apr, Gmr; mariner transposon delivery vector | S. Lory | |
| pPS1472 | Apr, Kmr; pCR2.1 with amplified PA1611 from PAO1 | This study | |
| pPS1476 | Apr; pEX18Ap-PA1611 (ligation of ∼2.0 kb blunt-ended EcoRI fragment of pPS1472 to PstI+EcoRI cleaved and blunt-ended pEX18Ap) | This study | |
| pPS1479 | Apr, Gmr; pEX18Ap-ΔPA1611::Gm (ligation of blunt-ended KpnI-NcoI fragment of pPS1476 to ∼1.1 kb SmaI Gmr::FRT fragment of pPS856) | This study | |
| pPS1503b | Apr, Gmr; pEX18Ap-ΔPA1627::Gm (ligation of the StuI-XhoI fragment of PCR-amplified ΔPA1627::Gm from PAO1 into the SmaI-SalI sites of pEX18Ap) | This study | |
| pPS1506b | Apr, Gmr; pEX18Ap-ΔPA1539::Gm (ligation of the MscI-XhoI fragment of PCR-amplified ΔPA1539::Gm into the SalI-SmaI sites of pEX18Ap) | This study | |
| pPS1505b | Apr, Gmr; pEX18Ap-ΔPA4890::Gm (ligation of the XhoI-SmaI fragment of PCR amplified ΔPA4890::Gm into SalI-SmaI sites of pEX18Ap | This study | |
| pPS1634 | Apr, Gmr; pUC18R6K-mini-Tn7T-Gm-pfabA::lacZY (ligation of three fragments, BamHI-XhoI fragment of pPS1633, ∼250 bp BamHI-EcoRV fragment of pPS790 and ∼6.2 kb SmaI-SalI fragment of pMC1403 into pPS1633) | This study | |
| pPS1633 | Apr, Gmr; pUC18R6K-mini-Tn7T-Gm | [37] | |
| pMC1403 | Apr; lacZY fusion cloning vector for construction of translational fusions | [17] | |
| pPS1733 | Apr, Kmr ; anr + (ligation of 0.8 kb PCR fragment amplified with Anr-UP-KpnI and Anr-DN-BamHI to pCR2.1 vector) | This study | |
| pPS1682 | Spra; anr + (ligation of 0.8 kb KpnI-BamHI fragment of pPS1733 into KpnI-BamHI cleaved pVLT35) | This study | |
| pPS1684 | Apr; anr + (ligation of 0.8 kb KpnI-BamHI fragment of pPS1733 into KpnI-BamHI cleaved pUCP20) | This study | |
| pPS1991b | Apr, Gmr; pPS1669 with attL1-pPA1612-attL2 to promote lacZ expression | This study | |
a
Abbreviations: Ap, ampicillin; att, λ attachment site (s); FRT, Flp recombinase target site; Gm, gentamycin; Km, kanamycin; MCS, multiple cloning site; p, promoters; Sp, spectinomycin
b
see text for plasmid or strain construction details.