TABLE 1.
Strains and plasmids
| Strain or plasmid | Characteristics | Reference or source |
|---|---|---|
| E. coli | ||
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac[F′ proAB lacIqZΔM15 Tn10(Tetr)] | Stratagene |
| DH5α | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 ΔlacU169(φ80 lacZΔM15) | Laboratory stock |
| A. tumefaciens | ||
| A348 | C58 background carrying pTiA6 | 13 |
| A348-3 | A348 with Kanr cartridge replacing a 4.6 kb KpnI fragment that contained virA on pTiA6 | 19 |
| UIA143 | autC58 recA::ery140 (pATC58) | 12 |
| A348-4 | A348-3 recA::ery140 | This study |
| Plasmids | ||
| pUC19 | E. coli cloning vector | New England Biolabs |
| pAW17 | pUC19 carrying virAH474Q at KpnI | This study |
| pAW18 | pUC19 carrying virA at KpnI | This study |
| pAW19 | pUC19 carrying Flag-tagged virA at KpnI | This study |
| pAW20 | pUC19 carrying virAE255L at KpnI | This study |
| pAW21 | pUC19 carrying virAL332E | This study |
| pYW10 | IncW E. coli/Agrobacterium shuttle vector pUCD2-derived (pSa origin of replication) | (4, 10, 40) |
| pAW10 | pYW10 with pUC19 cloning site at NcoI | This study |
| pAW13 | pAW10 carrying virAE255L | This study |
| pAW14 | pAW10 carrying virAH474Q | This study |
| pAW15 | pAW10 carrying virAH474Q and virAE255L | This study |
| pAW16 | pAW10 carrying wild type virA at KpnI | This study |
| pAW16a | pAW10 with Flag-tagged wild-type virA | This study |
| pAW50 | E. coli/Agrobacterium shuttle vector with pRi-derived origin of replication | This study |
| pAW52 | pAW50 carrying wild-type virA at KpnI | This study |
| pAW52a | pAW50 containing Flag-tagged virA at KpnI | This study |
| pAW53 | pAW50 containing virAE255L at KpnI | This study |
| pAW54 | pAW50 containing virAE255L and virAH474Q | This study |
| pAW55 | pAW50 containing virAH474Q at KpnI | This study |
| pAW56 | pAW50 containing virAL332E at KpnI | This study |
| pAW59 | pAW10 containing virAL332E at KpnI | This study |
| pLV2 | pAW10 containing virAE255L+H474Q at KpnI | This study |
| pYW47 | PN25-virG at Ncol site of pYW10 | 40 |
| pYW48 | pYW47 with wild-type virA at KpnI | 40 |
| pAW58 | pYW47 with virAH474Q at KpnI | This study |
| pAW60 | pAW47 with virAL332E at KpnI | This study |
| pAW61 | pYW47 with virAE255L at KpnI | |
| pSW209 | IncP plasmid carrying PvirB-lacZ fusion, Kanr | S. C. Winans |
| pSW209Ω | Derivative of pSW209 that is also Specr | 40 |