TABLE 1.
Bacterial strains and plasmids used in this study
| Bacterial strain or plasmid | Relevant genotype, phenotype, or characteristic(s)a | Reference or source |
|---|---|---|
| Escherichia coli | ||
| DH5α | supE44 ΔlacU169(φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | 42 |
| S17-1 | pro (r− m+) Mob+ Tpr Smr | 44 |
| Agrobacterium tumefaciens | ||
| C58 | Wild-type pathogenic strain carrying pTiC58 | Our collection |
| NT1 | Ti plasmid-cured C58 | 11 |
| NTL4 | NT1 derivative with tetA tetR mutation | 29 |
| Plasmids | ||
| pBluescript SK(±) | Cloning vector; Apr | Stratagene |
| pCF1 | 3,680-bp BamHI fragment 13 encoding traI, trbB, and repA from pTiC58 cloned in pTZ18U; Apr | 21 |
| pCMA1 | pTiC58traM::nptII; Kmr | 20 |
| pH13Km | 4,906-bp HindIII fragment containing the rep region of pTiC58 ligated with 3,484-bp HindIII fragment from Tn5; Smr Bmr Kmr | This study |
| pJB3 | Broad-host-range IncP cloning vector; Apr | 5 |
| pKP19 | traI::lacZY including 161 bp of the traI promoter region cloned into pLAFR6 | 21 |
| pLKC482 | pUC8-derived vectors for constructing translational fusion with lacZ; Apr Kmr | 48 |
| pPLH13 | 4,906-bp HindIII fragment containing the rep region of pTiC58 cloned in pBluescript SK(+); Apr | This study |
| pPLrep | 6.3-kb HindIII-HindIII fragment containing lacZY genes from pLKC482 cloned into SphI site in pCF1; Kmr Apr | This study |
| pPLrep4 | 6.7-kb PstI-PstI fragment from pPLrep cloned into pJB3; Apr | This study |
| pPLrep5 | 6.7-kb HindIII-HindIII fragment from pPLrep cloned into pJB3; Apr | This study |
| pPLrep6 | 5.6-kb BglII-BglII fragment from pPLrep cloned into pJB3; Apr | This study |
| pRK415 | Broad-host-range IncP cloning vector; Tcr | 24 |
| pRKE33 | 1.8-kb EcoRI fragment encoding traR from pTiC58 cloned into pRK415; Tcr | 36 |
| pSVB33 | 1.8-kb EcoRI fragment encoding traR from pTiC58 cloned into pSa152; Kmr Gmr Smr | 36 |
| pTB12 | traI::lacZY including 167 bp of the traI promoter region cloned in pLAFR6 | This study |
| pTiC58ΔaccR | A Trac mutant of pTiC58 | 4 |
a
Abbreviations: Smr, streptomycin resistance; Tcr, tetracycline resistance; Cbr, carbenicillin resistance; Kmr, kanamycin resistance; Apr, ampicillin resistance; Gmr, gentamicin resistance; Spr, spectinomycin resistance; Tpr, trimethoprim resistance; Bmr, bleomycin resistance.