TABLE 1.
Strains and plasmids
| Strain or plasmid | Characteristics | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| S17-1/λpir | RK2 tra regulon, pir negative, host for pir-dependent plasmid | 22 |
| DH5α | λ−φ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) supE44 thi-1 gyrA relA1 | GIBCO BRL |
| A. tumefaciens | ||
| A348 | C58 chromosomal background carrying pTiA6 | 18 |
| AB20 | A348 chvD gene was disrupted by insertion of the pZL101 suicide plasmid, Kanr | This study |
| AB21 | A348 “21-5” gene was disrupted by insertion of the pZL102 suicide plasmid, Kanr | This study |
| AB22 | A348 chvD gene was disrupted by insertion of the pZL103 suicide plasmid, Kanr | This study |
| S. cerevisiae EGY 48 | MATα trp1 his3 ura3 lexAops-LEU2 | 20 |
| Plasmids | ||
| pBSII | ColE1 Ampr, cloning vector | Stratagene |
| pJB20 | Broad-host-range plasmid, derivative of pUCD2; IncW Ampr Tetr | 5 |
| pJB93 | A 528-bp virB8 EcoRI fragment without a transmembrane domain in pJK202, HIS3 Ampr | This study |
| pJG4-5 (pV0) | Cloning prey vector for the construction of activator fusion, TRP1 Ampr | 20 |
| pJK202 | Cloning bait vector for the construction of LexA fusions, HIS3 Ampr | 20 |
| pMutA-G665D | IncRi derivative plasmid carrying virA (G665D) and wild-type virG, Ampr | 29 |
| pSH18-34 | Contains Gal1-LexAop-lacZ, a reporter gene in yeast, URA3 Ampr | 20 |
| pV1 | Modified version of pJG4-5 with EcoRI site carrying +1 reading frame, TRP1 Ampr | This study |
| pV2 | Modified version of pJG4-5 with EcoRI site carrying +2 reading frame, TRP1 Ampr | This study |
| pV01-1 | 835-bp Tsp509I fragment of A348 DNA containing a portion of chvD in pJG4-5, TRP1 Ampr | This study |
| pV21-5 | 468-bp Tsp509I fragment of A348 DNA in pV2, TRP1 Ampr | This study |
| pVK112 | R6K suicide vector, lacZY for transcriptional fusions, Kanr | 22 |
| pYW12 | pJB20 derivative, with lac promoter followed by the multiple cloning site from pUC19; Ampr | 39 |
| pYW15c | Broad-host-range expression vector, PN25-MCS-STOP, IncW and pBR322ori, Ampr | 39 |
| pYW48 | pYW15 derivative plasmid with PN25∷virG and wild-type virA, IncW Ampr | 39 |
| pZL35 | 2.7-kb EcoRV fragment containing the chvD gene from pZL78 cloned into pBSII, Ampr | This study |
| pZL39 | 2.7-kb EcoRV chvD-containing fragment from pZL78 cloned into pJB20 ScaI site, Tetr | This study |
| pZL78 | 7.8-kb HindIII fragment including chvD open reading frame cloned into pBSII, Ampr | This study |
| pZL81 | 2.7-kb PCR fragment containing chvD open reading frame and promoter cloned into pYW12 KpnI site; Ampr | This study |
| pZL82 | pZL81 derivative with mutation in Walker A1 ATP-binding site of chvD; Ampr | This study |
| pZL83 | pZL81 derivative with mutation in Walker A2 ATP-binding site of chvD; Ampr | This study |
| pZL84 | pZL81 derivative with mutations in both Walker A ATP-binding sites of chvD; Carr | This study |
| pZL101 | 449-bp chvD (nucleotides 335–784) PCR fragment from pV01-1 cloned into EcoRI site of pVK112 in the same orientation as lacZ; Kanr | This study |
| pZL102 | 398-bp v21-5 (nucleotides 28–426) PCR fragment from pV21-5 cloned into EcoRI site of pVK112; Kanr | This study |
| pZL103 | 449-bp chvD (nucleotides 335–784) PCR fragment from pV01-1 cloned into EcoRI site of pVK112 in the orientation opposite to that of lacZ; Kanr | This study |