Table 1.

Bacterial strains and plasmids used

Bacterial strain or plasmidRelevant genotype, phenotype, or characteristic(s)aSource or reference(s)
Escherichia coli
 DH5α supE44 ΔlacU169(φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 47
 S17-1 pro Res Mod+Mob+ Ampr Cmr TprStrr 15
 S17-1λ-pir pro ResMod+ Mob+ Ampr CmrTpr Strrλ::pir 24
 2174(pPH1JI) met pro GmrSpr, used to isolate homogenotes 24
Agrobacterium tumefaciens
 C58Wild-type pathogenic strain carrying pTiC58 and pAtC58Our collection
 NT1Ti plasmid-cured C58 15
 NT1(pTiC58ΔaccR)NT1 harboring a Tracmutant of pTiC58 6
 UIA5Ti plasmid- and cryptic plasmid-cured C58, RifrStrr 14
 C58C1RSTi plasmid-cured C58, RifrStrr 15
 C58C1ECTi plasmid-cured C58, Eryr Cmr Our collection
 pCF13,680-bp BamHI fragment 13 encoding traI, trbB, and repA from pTiC58 cloned in pTZ18U, Ampr 38
 pDCI41E33 traG::lacZ traR in pDSK519, Kmr, autoinducer reporter plasmid 15
 pDEK-m b pTiC58ΔaccR::Tn3HoHo1 marker-exchanged mutants, Cbr 42
 pDSK519Broad-host-range IncQ cloning vector, Kmr 41
 pHM1Delivery plasmid for mini-Tn5Ptrb, AmprKmr This study
 pHM25 HindIII fragment 8 containing Tn3HoHo1 from pPLE2-25 cloned in pRK415, Tcr 38
 pHM25-70pHM25::mini-Tn5Ptrb, Tcr Kmr This study
 pJB3Broad-host-range IncP cloning vector, Ampr 9
 pKK38Broad-host-range IncP cloning vector, tacpromoter, Tcr David Nunn
 pKK38ASHDerivative of pKK38 with extra cloning sites, Tcr This study
 pPLE2-25pPLE2 with Tn3HoHo1 insertion intrbE, Cbr 38, 42
 pPLKmPtrbKm-Ptrb cassette from mini-Tn5Ptrb cloned in pBluescript SK(+), Ampr Kmr This study
 pPLK-n c pTiC58ΔaccR::mini-Tn5Ptrbmarker-exchanged mutants, Kmr This study
 pPLKΔCpTiC58ΔaccR trbC::Km-Ptrb marker-exchanged mutant, Kmr This study
 pPLKΔKpTiC58ΔaccR trbK::nptI marker-exchanged mutant, Kmr This study
 pPLtra tra region andtraR from pTiC58 cloned in pDSK519, Tcr This study
 pPLtrbX d PCR-generated individual trb gene ORFs cloned in pKK38ASH or pKK38, Tcr This study
 pRK415Broad-host-range IncP cloning vector, Tcr 41
 pRKtrb11,003-bp BglII-XbaI fragment containing the trb region of pTiC58 cloned in pRK415, Tcr This study
 pRKtrb-n pRKtrb::mini-Tn5Ptrbmutants, Tcr Kmr This study
 pRKtrbΔCpRKtrb with a Km-Ptrb cassette intrbC, Tcr Kmr This study
 pSB315pUC4K derivative containing nptI, Kmr 32
 pSVB331.8-kbEcoRI fragment encoding traR from pTiC58 cloned in pSa152, Kmr Gmr 46
 pTra17-52pTiC58ΔaccR nocR::Tn5 17-52, NocTrac Kmr 6
 pUTmini-Tn5KmDelivery plasmid for mini-Tn5Km, AmprKmr 20
  • a Abbreviations: Rifr, rifampin resistance; Strr, streptomycin resistance; Tcr, tetracycline resistance; Cbr, carbenicillin resistance; Kmr, kanamycin resistance; Ampr, ampicillin resistance; Cmr, chloramphenicol resistance; Gmr, gentamicin resistance; Emr, erythromycin resistance; Spr, spectinomycin resistance; Tpr, trimethoprim resistance; Trac, transfer constitutive.

  • b m refers to the allele number of the Tn3HoHo1 insertion as shown in Fig. 3C.

  • c n refers to the allele number of the mini-Tn5Ptrb insertion as shown in Fig. 3B.

  • d trbX refers to trbgenes. X can be B, C, D,E, J, K, L, F,G, H, or I.