TABLE 1.
Strains and plasmids used in this study
| Strain | Characteristics | Source or reference |
|---|---|---|
| A. tumefaciens strains | ||
| C58 | Wild-type pathogenic strain carrying pTiC58 | Our collection |
| NT1 | Derivative of C58, cured of pTiC58 | 53 |
| NTL4 | Derivative of C58, cured of pTiC58; ΔtetRS | 31 |
| C58C1RS | Derivative of C58, cured of pTiC58; Rifr StrR | Our collection |
| E. coli strains | ||
| MG1655 | K-12 derivative; F− λ−ilvG rfb-50 rph-1 | J. Cronan, Jr. |
| DH5α | supE44 ΔlacU169(φlacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Invitrogen |
| WM5979 | lacIqrrnB3 ΔlacZ4787 hsdR514 Δ(araBAD)567 Δ(rhaBAD)568 rph-1 Δatt λ::pAE12-Δ(oriR6K/cat::frt5) Δ4229(dapA)::frtΔ(endA)::frt attHK::pJK1006::Δ1/2 (ΔoriR6K-cat::frt5 ΔtrfA::frt) | W. Metcalf |
| Other strains | ||
| Rhizobium leguminosarum bv. viciae 3841 | Derivative of R. leguminosarum bv. viciae strain 300; Smr | J. A. Downie |
| Sinorhizobium meliloti 1021 | Derivative of wild-type strain SU47; Smr | S. R. Long |
| Caulobacter crescentus CB15 | Wild-type strain | J. M. Slauch |
| Pseudomonas fluorescens 1855-344 | Wild-type strain | Our collection |
| Brucella abortus 2308 | Wild-type, smooth, virulent laboratory strain | R. M. Roop II |
| Ralstonia solanacearum AW1 | Wild-type strain | T. P. Denny |
| Plasmids | ||
| pTiC58 | Wild-type Ti plasmid of strain C58; Noc+ Vir+ Trai | Our collection |
| pKPC12 | pTiC58 (traR::lacZ) TraR−; Noc+ Vir+ Tra− Kmr | 41 |
| pBluescript II SK(+) | Cloning vector; Co1E1 f1(+) Ampr | Stratagene |
| pBBR1MCS-2 | Broad-host-range vector; Kmr | 26 |
| pBBR1MCS-3 | Broad-host-range vector; Tetr | 26 |
| pBBR1MCS-5 | Broad-host-range vector; Gmr | 26 |
| pZLQ | pBBR1MCS2-derived expression vector containing trc promoter; Kmr | 30 |
| pZLQR | pZLQ containing C58 traR under the control of Ptrc | 30 |
| pSRKGm(α−) | pBBR1MCS-5 derivative; ΔlacZα | This study |
| pSRKKm(α−) | pBBR1MCS-2 derivative; ΔlacZα | This study |
| pSRKTc(α−) | pBBR1MCS-3 derivative; ΔlacZα | This study |
| pSRKGm(lacI) | pSRKGm(α−) containing lacIq and placZ | This study |
| pSRKGm | pBBR1MCS-5-derived broad-host-range expression vector containing lac promoter and lacIq, lacZα+, and Gmr | This study |
| pSRKKm | pBBR1MCS-2-derived broad-host-range expression vector containing lac promoter and lacIq, lacZα+, and Kmr | This study |
| pSRKTc | pBBR1MCS-3-derived broad-host-range expression vector containing lac promoter and lacIq, lacZα+, and Tetr | This study |
| pSRKKm::uidA | pSRKKm containing placZ::uidA | This study |
| pSRKGmtraR | pSRKGm containing traR from pTiC58 inserted between NdeI and BamHI sites | This study |
| pZLB251 | pRG970b containing traA to traC intergenic region and traA::lacZ fusion | 30 |
| pYZ1 | pRK415 containing BamHI fragment 24 of pTiC58, source of traM gene | 21 |
| pGPO3 | Derivative of pDSK519 containing traR from pTiC58 under the control of Plac; lacIq Kanr | 17 |
| pGPO3-I-gnt | pGPO3 containing lacIq and gentamicin resistance cassette Gmr; Kanr | This study |
| pKK38-I | Expression vector containing Ptrc and lacIq; Tetr | 32 |
| pKKTR2-I | pKK38-I expressing traR from pTiC58 under the control of Ptrc; lacIq Tetr | 32 |