TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| Agrobacterium strains | ||
| A348 | A136(pTiA6) | 21 |
| A6007 | A348 Pho− Smr | 11 |
| A6880 | A6007 chvH::TnphoA | 12 |
| At13000 | A348 ΔchvH::Ω fragment, contains pWT187kan | This study |
| At13001 | At13000 without pWT187kan | This study |
| E. coli strains | ||
| DH5α | endA1 hsdR17 supE44 thi-1 recA1 gyrA96 relA1 Δ(argF-lacZYA)U169 φ80dlacZΔM15 | Gibco-BRL |
| TG1 | supE hsdΔ5 thi Δ(lac-proAB) F′ (traD36 proAB+ lacIqlacZΔM15) | U.S. Biochemical Corp. |
| Plasmids | ||
| pACL2 | pRK7813 cosmid clone which complements A6880 | 12 |
| pSW172 | IncP broad-host-range vector with lac promoter | 14 |
| pSW213 | IncP broad-host-range vector with lac promoter and lacIq | 14 |
| pTC110 | pUCD2 ΔPvuII-EcoRV | 13 |
| pBBR1MCS-4 | Broad-host-range vector | 32 |
| pUC18 | Multicopy vector | 55 |
| pUC19Ω | SmaI Ω fragment in pUC19 | 10 |
| pPR1068 | pMAL-c2 derivative; NdeI at the start of MalE | New England Biolabs |
| pSP329 | pTJS75 derivative with α-complementation group and multicloning site from pUC18, IncP, Tcr | 11 |
| pSP329Gm | pSP329 derivative, IncP, Gmr | D. M. Raineri |
| pJQ200SK | sacB suicide vector | 44 |
| pUCD2 | IncW broad-host-range vector | 16 |
| pBSIIKS+.NdeI | pBluescriptIIKS+ derivative; NdeI at the start of lacZ′ | 9 |
| pAB2002 | Vector for transcriptional fusion with lacZ | 7 |
| pTC111 | virE2::lacZ translational fusion in pTC110 | T. C. Charles, unpublished |
| pTC112 | virB1::lacZ translational fusion in pTC110 | T. C. Charles, unpublished |
| pUFR047 | IncW broad-host-range vector | 17 |
| pPR1068-E1+E2 | Ptac-virE operon in pPR1068 | 18 |
| pUFR047-E1+E2 | Ptac-virE operon in pUFR047 | 18 |
| pSL59 | virA-phoA translational fusion | 19 |
| pTC234 | 7.4-kb KpnI-SacI fragment from pACL2 in pSP329 | This study |
| pTC234Ω9A::Tn5-B20 | Tn5-B20 insertion into chvH gene of pTC234 | This study |
| pWT131 | 4.4-kb KpnI-BamHI fragment in pBluescriptIISK+ | This study |
| pWT142 | pUC18 carrying a 7.6-kb SalI fragment from A6880, Kmr | This study |
| pWT151 | 2.0-kb EcoRI fragment of pTC234 in pSW172 | This study |
| pWT154 | Ptac-chvH in pPR1068 | This study |
| pWT155 | Ptac-chvH in pUCD2 | This study |
| pWT158 | 2-kb HpaI-ClaI fragment of pPR1068-E1+E2 in pBBR1MCS-4 | This study |
| pWT159 | Ptac-virE1 virE2-lacZ transcriptional fusion in pBBR1MCS-4 | This study |
| pWT160 | virG-lacZ translational fusion in pTC110 | This study |
| pWT179 | Ptac-driven E. coli efp in pPR1068 | This study |
| pWT181 | Ptac-driven E. coli efp in pSP329Gm | This study |
| pWT183 | Plac-chvH in pBSIIKS+.NdeI | This study |
| pWT187 | 1.6-kb PvuII fragment containing Plac-chvH in pSW213 | This study |
| pWT187kan | pWT187 plus a kanamycin resistance cassette | This study |
| pWT188 | 445-bp chvH coding region replaced by Ω in pWT183 | This study |
| pWT191 | pWT131 with the 1.29-kb NdeI-EcoRV fragment replaced by 2.8-kb NdeI-EcoRV fragment from pWT188 | This study |
| pWT193 | 6-kb KpnI-SstI fragment from pWT191 in pJQ200SK | This study |