TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype and/or phenotypea | Source or reference |
|---|---|---|
| Strains | ||
| A. caviae | ||
| Sch3 | Clinical isolate | 54 |
| Sch3N | Sch3, spontaneous Nalr | This study |
| AAR269 | flaA::Kmr | This study |
| AAR27 | flaB::Kmr | This study |
| AAR31 | flaA::Cmr, flaB::Kmr | This study |
| AAR150 | flaG::Kmr | This study |
| AAR59 | flaH::Kmr | This study |
| AAR8 | flaJ::Kmr | This study |
| E. coli | ||
| CC118λpir | Δ(ara leu)7697 araD139 ΔlacZX74 glaE glaK phoA20 thi-1 rspE rpoB(Rfr) argE(Am) recA1αpir+ | 17 |
| S17-1λpir | hsdR, pro, recA, RP4-2 in chromosome Km::Tn7 (Tc::Mu) λpir, Tpr Smr | 41 |
| XL1-Blue | endA1 recA1 hsdR17 supE44 thi-1 gyrA96 relA1 lac [F′ proAB laclqZΔM15 Tn10 (Tcr)] | Stratagene |
| Plasmids | ||
| pUC19 | High-copy-number cloning vector, MCS, Ampr | Gibco-BRL |
| pUTmini-Tn5Cm | bla, ori R6K, mob RP4, tnp gene of Tn5-IS50r that lacks NotI site, MCS of M13tg131; 8.7 kb, Apr Cmr | 9 |
| pUC4-KIXX | Source of Tn5-derived nptII gene (Kmr) | Pharmacia |
| pBBR1MCS | Broad-host-range vector; IncP, W, and -Q; Col E1; and p15A compatible, containing pBluescript II (KS)- lacZα-polylinker, Cmr | 32 |
| pBBR1MCS-3 | Broad-host-range vector; IncP, -W, and -Q; Col E1; and p15A compatible, carrying Tcr | 31 |
| pKNG101 | ori R6K, mob RK2, strAB sacBR, 6.8 kb; Smr | 21 |
| pARP1 | pUC19 with a 900-bp fla PCR insert | This study |
| pARP2 | pBBR1MCS with a 3.2-kb SacI insert of A. caviae Sch3N DNA | This study |
| pARP126 | pBBR1MCS with a 3.6-kb SacI insert of A. caviae Sch3N DNA | This study |
| pARA100 | 1.5-kb XbaI-SmaI fragment containing flaA from pARP2, inserted at the pKNG101 | This study |
| pARA110 | 1.4-kb Kmr cassette from pUC4-KIXX inserted at the PvuII site of flaA in pARA100 | This study |
| pARA120 | 3.7-kb end-filled HindIII fragment from pUTmini-Tn5Cm, inserted at the PvuII site in pARA100 | This study |
| pARB200 | 1.2-kb SacI-HincII fragment containing flaB from pARP126 in pUC19 | This study |
| pARB210 | 1.8-kb KmrBamHI-SmaI fragment from pUC4-KIXX inserted at the BglII-EcoRV sites of flaB in pARB200 | This study |
| pARB220 | 3.7-kb CmrEcoRI fragment of pUTmini-Tn5Cm inserted at the EcoRI site of pARB210 | This study |
| pARB230 | 2.2-kb BamHI fragment from pARB220 inserted at the BamHI site of pKNG101 | This study |
| pARP400 | 3.6-kb SacI fragment from pARP126 inserted into pUC19 | This study |
| pARG410 | 1.4-kb KmrSmaI fragment from pUC4KIXX inserted at the StuI site of flaG in pARP400 | This study |
| pARG420 | 4.2-kb SalI fragment from pARG410 inserted at the SalI site of pKNG101 | This study |
| pARH500 | 2.2-kb SacI-StuI fragment from pARP400 inserted at SacI-SmaI site on pUC19. | This study |
| pARH510 | 1.4-kb KmrSmaI fragment from pUC4KIXX inserted at the SmaI site of flaH pARH500 | This study |
| pARH520 | 3.6-kb SacI-BamHI fragment from pARH510 inserted at SacI-BamHI sites of pKNG101 | This study |
| pARH530 | 3.7-kb CmrBamHI fragment from pUTmini-Tn5Cm inserted at the BamHI site of pARH520 | This study |
| pARJ600 | 1.4-kb KmrSmaI fragment from pUC4KIXX inserted at BalI site of flaJ in pARH500 | This study |
| pARJ610 | 3.6-kb SacI-BamHI fragment from pARJ600 inserted at the SacI-BamHI sites of pKNG101 | This study |
| pARJ620 | 3.7-kb CmrBamHI fragment from pUTmini-Tn5Cm inserted at the BamHI site of pARJ610 | This study |
| pARP1264 | 1.8-kb NaeI-SacI fragment from pARP2 inserted at the SmaI-SacI site of pBBR1MCS-3 | This study |
| pARP1265 | 3.6-kb SacI insert from pARP126 inserted in the SacI site of pBBR1MCS-3 | This study |
| pARP1266 | 3.6-kb SacI insert from pARP126 inserted in the SacI site of pARP1264 | This study |
a
Phenotype abbreviations are as follows: resistance (r); Amp (ampicillin); Cm (CHL); Km (KAN); Nal (NAL); Sm (STR); and Tc (tetracycline). MCS, multiple cloning site.