TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Source or reference |
|---|---|---|
| A. tumefaciens | ||
| A6 | Wild-type octopine strain | A. Kerr |
| A6(traMk588) | A6 with traM replaced with traM of strain K588, which is dysfunctional | 25 |
| A6(traMk588 ΔtraM2) | A6(traMk588) carrying in-frame deletion of traM2 | 25 |
| A6(traMk588 ΔtraM2; traR2::Tn5) | A6(traMk588 ΔtraM2) carrying Tn5 insertion of traR2, Kanr | This study |
| A6(traMk588 pDSK-traR2) | A6(traMk588) with overexpression of traR2, Kanr | This study |
| NT1(pDSK-traR pLA-traG::lacZ) | Also named NT1(traR tra::lacZ749), AHL signal reporter strain, Kanr Tcr | 9 |
| A6(traMk588 pLA-traG::lacZ) | A6(traMk588) carrying AHL-responsive reporter plasmid pLA-traG::lacZ, Tcr | This study |
| A6(traMk588 ΔtraM2 pLA-traG::lacZ) | A6(traMk588 ΔtraM2) carrying pLA-traG::lacZ vector, Tcr | This study |
| A6(traMk588 ΔtraM2 traR2::Tn5 pLA-traG::lacZ) | A6(traMk588 ΔtraM2; traR2::Tn5) carrying pLA-traG::lacZ vector, Kanr Tcr | This study |
| A6(traMk588 pDSK-traR2 pLA-traG::lacZ) | A6(traMk588, pDSK-traR2) carrying pLA-traG::lacZ vector, Kanr Tcr | This study |
| C58C1 | Derivative of nopaline strain C58 cured of Ti plasmid used for conjugation analysis, Rifr | S. Q. Pan |
| Ach5C3 | Derivative of wild-type octopine strain Ach5, Ti plasmid cured, Rifr | S. K. Farrand |
| Ach5C3(pLA-traG::lacZ) | Ach5C3 carrying pLA-traG::lacZ vector, Tcr | This study |
| Ach5C3(pLA-traG::lacZ pDSK-traR2) | Ach5C3(pLA-traG::lacZ) carrying pDSK-traR2 vector, Kanr Tcr | This study |
| A6(traMk588, ΔtraI) | A6(traMk588) with in-frame deletion of traI gene | This study |
| A6(traMk588 ΔtraI traI2::Tn5) | A6(traMk588, ΔtraI) with Tn5 insertion of traI2, Knar | This study |
| A6(traMk588 ΔtraI traI2::Tn5 pLA-traI2) | A6(traMk588 ΔtraI traI2::Tn5) carrying plasmid pLA-traI2, Knar Tcr | This study |
| A6(traMk588 ΔtraI traI2::Tn5 pLA-traI) | A6(traMk588 ΔtraI traI2::Tn5) carrying plasmid pLA-traI, Knar Tcr | This study |
| C1A6MR1 | Transconjugant of A6(traMk588 ΔtraM2; traR2::Tn5) and C58C1, Rifr Kanr | This study |
| C1A6MR2 | Transconjugant of A6(traMk588 ΔtraM2; traR2::Tn5) and C58C1, Rifr Kanr | This study |
| C1A6MR3 | Transconjugant of A6(traMk588 ΔtraM2; traR2::Tn5) and C58C1, Rifr Kanr | This study |
| C58C1(pTiA6) | C58C1 carrying Ti plasmid of A6 | 25 |
| E. coli | ||
| DH5α (λpir) | supE44 ΔlacU169(ϕ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 λpir | Laboratory collection |
| BW020767(pRL27) | Harbors Tn5 for A. tumefaciens mutagenesis | 49 |
| Plasmids | ||
| pK18mobsacB | Broad-host-range gene replacement vector, Sucr Kanr | 50 |
| pK18-traI | pK18mobsacB harboring deleted traI-flanking region, Kanr | This study |
| pLAFR3 | IncP, broad-host-range cosmid vector, Tcr | 51 |
| pLA-traG::lacZ | pLAR3 carrying traG gene transcriptionally fused with promoterless lacZ, Tcr | 9 |
| pLA-traI | pLAFR3 harboring traI gene from A6, Tcr | This study |
| pLA-traI2 | pLAFR3 harboring traI2 gene from A6, Tcr | This study |
| pDSK | IncQ, broad-host-range plasmid, Kanr | 52 |
| pDSK-traR | pDSK harboring traR gene from A6, Kanr | 9 |
| pDSK-traR2 | pDSK harboring traR2 gene from A6, Kanr | This study |
a
Abbreviations: Ampr, ampicillin resistant; Kanr, kanamycin resistant; Tcr, tetracycline resistant; Rifr, rifampincin resistant; Sucr, sucrose resistant.