TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant characteristic(s) | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac[F′ proAB lacIqZΔM15 Tn10 (Tcr)] | Stratagene |
| DH5α | recA1 endA1 hsdR17 supE44 gyrA96 relA1Δ(lacZYA-argF)U169 (φ80dlacZΔM15) | Invitrogen |
| A. tumefaciens | ||
| A348 | A136 containing pTiA6NC | 19 |
| A136 | Strain C58 cured of pTi plasmid | 48 |
| DL8 | A136 chvE deletion, Kmr | This study |
| Plasmids | ||
| pBluescript II KS(+/−) | Cloning vector, ColE1 Apr | Stratagene |
| pJZ4 | 5.1-kb fragment containing PvirB-lacZ-nptII cloned into pMON596, Inc P Specr | 53 |
| pRG109 | PN25-His6-virG in pJZ4, Specr | This study |
| pRG108 | PN25-His6-virG(N54D) in pJZ4, Specr | This study |
| pYW47 | PN25-His6-virG in pYW15, IncW Apr | 47 |
| pAM13 | PN25-His6-virG(N54D) in pYW15, IncW Apr | 32 |
| pSM102 | occQ::lacZ, IncP Apr | 39 |
| pVRA8 | virA from pTiA6 in pUCD2, pBR322ori, IncW Apr | 25 |
| pMutA | virA(G665D) in pUCD2, pBR322ori, IncW Apr | 29 |
| pYW15 | Broad-host-range expression vector, IncW Apr | 47 |
| pFF4 | PN25-virA in pQE50, ColE1 Apr | This study |
| pFQ25 | PN25-virA and PN25-virG, IncW Apr | This study |
| pYW21 | PN25-His6-virA(285-829) in pYW15, Apr | 45 |
| pYW39 | PN25-His6-virA(285-829)(G665D) in pYW15, Apr | 47 |
| pAM23 | PN25-His6-virA(285-711) in pYW15, Apr | 32 |
| pAM28 | PN25-His6-virA(285-711) (G665D) in pYW15, Apr | 32 |
| pRG100 | PN25-His6-virA(426-829) (G665D) in pYW15, Apr | This study |
| pRG118 | PN25-His6-virA(426-711) (G665D) in pYW15, Apr | This study |
| pRG119 | PN25-His6-virA(426-711) in pYW15, Apr | This study |
| pRG120 | PN25-His6-virA(426-829) in pYW15, Apr | This study |
| pRG135 | virA(1-711) in pYW15, Apr | This study |
| pRG157 | chvE from A136 in pUC19, ColE1 Apr | This study |
| pRG159 | Kmr gene cloned in BamH I site of chvE in pRG157, ColE1 Apr Kmr | This study |
| pRG162 | virA(E210V) in pBluescript II KS (+/−), ColE1 Apr | This study |
| pRG165 | PN25-chvE in pYW15, Apr | This study |
| pRG166 | PN25-chvE(T211M) in pYW15, Apr | This study |
| pRG168 | virA and PN25-chvE in pYW15, Apr | This study |
| pRG169 | virA and PN25-chvE(T211M) in pYW15, Apr | This study |
| pRG170 | virA(E210V) and PN25-chvE in pYW15, Apr | This study |
| pRG171 | virA(E210V) and PN25-chvE(T211M) in pYW15, Apr | This study |