TABLE 1.
Bacterial strains and plasmids used in this study
| Bacterial strains and plasmids | Genotype or comments | Source or reference |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| CC118-λpir | Δ(ara-leu)7967 araD139 ΔlacX74 galE galK ΔphoA20 thi-1 rpsE rpoB argE(Am) recA1 | Laboratory collection |
| DH5a-λpir | DlacU169 (F80lacZDM15) recA1 endA1 hsdR17 thi-1 gyrA96 relA1 λpir phage lysogen | Laboratory collection |
| SY327-λpir | (lac pro) argE (Am) rif nalA recA56 (λpir) | Laboratory collection |
| Klebsiella pneumoniae | ||
| Kp52145 | Clinical isolate; serotype O1:K2; RifR | (3, 28) |
| 52145-ΔwcaK2 | Kp52145, ΔwcaK2, capsule mutant; RifR | (32) |
| 52OmpA2 | Kp52145, ΔompA::Cm, ompA gene is inactivated; RifR, CmR | (37) |
| 52ΔwcaK2OmpA | 52145-ΔwcaK2, ΔompA::Cm, capsule mutant in which ompA is inactivated; RifR, CmR | (37) |
| 52ΔwaaL | Kp52145, ΔwaaL, waaL is inactivated; nonpolar mutant; RifR | (68) |
| 52ΔwabM | Kp52145, ΔwabM, wabM is inactivated; nonpolar mutant; RifR | (50) |
| 52ΔwabH | Kp52145, ΔwabH, wabH is inactivated; nonpolar mutant; RifR | (50) |
| 52ΔwabK | Kp52145, ΔwabK, wabK is inactivated; non polar mutant; RifR | (50) |
| 52145-ΔwcaK2ΔwaaL | 52145-ΔwcaK2; ΔwaaL, waaL is inactivated; non polar mutant; RifR | (6) |
| 52145-ΔwcaK2ΔwabM | 52145-ΔwcaK2; ΔwabM, wabM is inactivated; non polar mutant; RifR | (6) |
| 52145-ΔwcaK2ΔwabK | 52145-ΔwcaK2; ΔwabK, wabK is inactivated; non polar mutant; RifR | (6) |
| 52145-ΔwcaK2ΔwabH | 52145-ΔwcaK2; ΔwabH, wabH is inactivated; non polar mutant; RifR | (6) |
| 52waaL::tn5 | Kp52145; ΔwaaL::tn5, waaL is inactivated; RifR, KmR | This study |
| 52entF::tn5 | Kp52145; entF::tn5, entF is inactivated; RifR, KmR | This study |
| 52145-ΔwcaK2entF | 52145-ΔwcaK2, entF:: pKNOCK-Cm_entF, capsule mutant in which entF is inactivated; RifR, CmR | This study |
| 52pulC::tn5 | Kp52145; ΔpulC::tn5, pulC is inactivated; RifR, KmR | This study |
| 52irp1 | Kp52145, irp1:: pKNOCK-Cm_IRP1, irp1 is inactivated; RifR, CmR | This study |
| 52iutA | Kp52145, iutA:: pKNOCK-Cm_iutA, iutA is inactivated; RifR, CmR | This study |
| 52glf | Kp52145, glf:: pSFglf, glf is inactivated; RifR, KmR | This study |
| 52ΔpulA | Kp52145, pulA:: pKNOCK-Cm_pulA, pulA is inactivated; RifR, CmR | This study |
| 52pulACom | 52ΔpulA harboring mini-Tn7TKmKpnpulA; PulA levels restored; RifR, CmR, KmR | This study |
| 52145-ΔwcaK2pulA | 52145-ΔwcaK2, pulA:: pKNOCK-Cm_pulA, capsule mutant in which pulA is inactivated; RifR, CmR | This study |
| 52145-ΔwcaK2pulACom | 52145-ΔwcaK2pulA mutant harboring mini-Tn7TKmKpnpulA; PulA levels restored; RifR, CmR, KmR | This study |
| 52ΔwaaLpulA | 52ΔwaaL, pulA:: pKNOCK-Cm_pulA, waaL mutant in which pulA is inactivated; RifR, CmR | This study |
| 52145-ΔwcaK2ΔwaaL pulA | 52145-ΔwcaK2ΔwaaL, pulA:: pKNOCK-Cm_pulA, capsule and waaL mutant in which pulA is inactivated; RifR, CmR | This study |
| Plasmids | ||
| pRL27 | Tn5 transposon delivery vector, KmR | (30) |
| pGEM-T Easy | Cloning plasmid, AmpR | Promega |
| pKNOCK-Cm | Suicide vector, R6K replication origin, CmR | (35) |
| pKNOCK-Cm_entF | pKNOCK-Cm containing an internal fragment of entF gene for mutant construction, CmR | This study |
| pKNOCK-Cm_IRP1 | pKNOCK-Cm containing an internal fragment of irp1 gene for mutant construction, CmR | This study |
| pKNOCK-Cm_iutA | pKNOCK-Cm containing an internal fragment of iutA gene for mutant construction, CmR | This study |
| pKNOCK-Cm_pulA | pKNOCK-Cm containing an internal fragment of pulA gene for mutant construction, CmR | This study |
| pSF100 | Suicide vector, R6K replication origin, KmR | (36) |
| pSFglf | pSF100 containing an internal fragment of glf gene for mutant construction, KmR | This study |
| pUC18R6KT-mini-Tn7TKm | pUC18R6KT-mini-Tn7TKm containing a Km cassette, AmpR, KmR | (37) |
| pTSNSK-Tp | pTSNSK-Tp containing a transposase for Tn7 insertion, KmR, TpR | (38) |
| pUC18R6KT-mini-Tn7TKm_pulACom | pUC18R6KT-mini-Tn7TKm containing pulA gene for complementation, AmpR, KmR | This study |
| p34S-Tp | Source of Tp cassette; AmpR, TpR | (40) |
| pPROBE′-gfp[LVA] | Plasmid containing gfp without promoter as reporter gene, KmR | (41) |
| pPROBE′-gfp[LVA]Tp | Trimethoprim resistance cassette cloned into SphI site of pPROBE′-gfp[LVA], TpR | This study |
| pPROBE′-gfp[LVA]Tp_entC | pPROBE′-gfp[LVA]Tp containing entC promoter region, TpR | This study |
| pPROBE′-gfp[LVA]Tp_ybtA | pPROBE′-gfp[LVA]Tp containing ybtA promoter region, TpR | This study |
| pPROBE′-gfp[LVA]Tp_psn | pPROBE′-gfp[LVA]Tp containing psn promoter region, TpR | This study |
| pPROBE′-gfp[LVA]Tp_iucA | pPROBE′-gfp[LVA]Tp containing iucA promoter region, TpR | This study |
| pGPLKpnCps | pGPL01 containing cps promoter region; AmpR | (37) |