TABLE 1.
Strains and plasmids
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| E. coli | ||
| DH5α | recA1 endA1 hsdR17(rK− mK+) supE44 thi-1 gyrA relA1 λ− φ80d/lacZΔM15 (ΔlacZYA-argF)U169 | Laboratory stock |
| A. tumefaciens | ||
| A136 | C58 heat cured of pTiC58 | Laboratory stock |
| A348 | A136 containing pTiA6 | Laboratory stock |
| A348ΔB8 | A348 containing a deletion in pTiA6 virB8 | 5 |
| Saccharomyces cerevisiae | ||
| EGY48 | ura3 his3 trp1 LexAop-leu2 | 16 |
| AD842 | EGY48 containing pSH18-34 | 11 |
| Plasmids | ||
| pEG202 | Cloning vector for the construction of LexA fusions | 16 |
| pJK202 | Derivative of pEG202 with a nuclear localization signal | A. Peterson |
| pJG4-5 | Cloning vector for the construction of activator fusions | 16 |
| pSH18-34 | A plasmid containing Gal1-LexAop-lacZ, a reporter gene in yeast | 16 |
| pAD1483 | 0.25-kb EcoRI fragment containing virB7 obtained by PCR amplification in pEG202 (activator-VirB7 A11 fusion; bp 6261–6509 of Ward et al. [26]) | This study |
| pAD1522 | 1.0-kb EcoRI fragment obtained by PCR amplification in pJK202 (LexA-VirB10 I47 fusion; bp 8111–9108) | This study |
| pAD1523 | 0.6-kb EcoRI fragment obtained by PCR amplification in pJK202 (LexA-VirB8 T60 fusion; bp 6562–7197) | This study |
| pAD1663 | 0.6-kb EcoRI-XhoI fragment of virB8S87L obtained by PCR amplification in pJK202 (LexA-VirB8S87L fusion at T60) | This study |
| pAD1526 | 0.9-kb EcoRI fragment obtained by PCR amplification in pJK202 (LexA-VirB9 T17 fusion; bp 7143–8019) | This study |
| pAD1447 | 0.25-kb EcoRI fragment containing virB7 obtained by PCR amplification of pAD1399 in pJG4-5 (activator-VirB7 A11 fusion) | 11 |
| pAD1493 | 1.0-kb EcoRI fragment obtained by PCR amplification in pJG4-5 (activator-VirB10 I47 fusion; bp 8111–9108) | This study |
| pAD1516 | 0.6-kb EcoRI-XhoI fragment obtained by PCR amplification in pJG4-5 (activator-VirB8 T60 fusion; bp 6562–7197) | This study |
| pAD1517 | 0.9-kb EcoRI fragment obtained by PCR amplification in pJG4-5 (activator-VirB9 T17 fusion; bp 7143–8019) | This study |
| pAD1529 | 0.35-kb EcoRI fragment obtained by PCR amplification in pJK202 (LexA-VirB8 T60-T172 fusion; bp 6562–6900) | This study |
| pAD1530 | 0.33-kb EcoRI fragment obtained by PCR amplification in pJK202 (LexA-VirB8 P145-P237 fusion; bp 6817–7098) | This study |
| pAD1531 | 0.33-kb EcoRI fragment obtained by PCR amplification in pJK202 (LexA-VirB9 T17-H122 fusion; bp 7143–7460) | This study |
| pAD1533 | 0.45-kb EcoRI fragment obtained by PCR amplification in pJK202 (LexA-VirB9 D75-P220 fusion; bp 7317–7754) | This study |
| pAD1492 | 0.4-kb EcoRI fragment obtained by PCR amplification in pJK202 (LexA-VirB9 L173-293 fusion; bp 7611–8019) | This study |
| pAD1528 | 0.7-kb EcoRI-SalI fragment obtained by PCR amplification in pJK202 (LexA-VirB10 I47-V277 fusion; bp 8111–8803) | This study |
| pAD1534 | 0.65-kb EcoRI fragment obtained by PCR amplification in pJK202 (LexA-VirB10 R161-377 fusion; bp 8453–9108) | This study |
| pAD1598 | 1.0-kb EcoRI fragment of pAD1493 in pRsetB (His-VirB10 fusion) | This study |
| pAD1599 | 1.0-kb EcoRI fragment of pAD1493 in pGEX1λT (GST-VirB10 fusion) | This study |
| pAD1601 | 0.6-kb EcoRI fragment of pAD1523 in pGEX1λT (GST-VirB8 fusion) | This study |
| pAD1606 | 0.9-kb EcoRI fragment of pAD1517 in pGEX1λT (GST-VirB9 fusion) | This study |