TABLE 1.
Bacterial strains and plasmids used for this study
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5αF′ | F′ endA1 hsdR17 recA1 gyrA Δ(lacZYA-argF)U169 φ80dlacZΔM15 | Laboratory stock |
| CJ236 | FΔ(HindIII)::cat ung-1 dut-1 | Laboratory stock |
| Agrobacterium tumefaciens | ||
| C58 | Harbors nopaline Ti plasmid pTiC58 | Laboratory stock |
| A136 | C58 cured of pTiC58 | Laboratory stock |
| A348 | A136 with octopine Ti plasmid pTiA6NC | Laboratory stock |
| A281 | A136 with succinamopine Ti plasmid pTiBo542 | Laboratory stock |
| AD802 | phoA derivative of A136 | 15 |
| A348ΔB | A348 with a deletion of pTiA6NC virB operon (PC1001 to PC1011) | 2 |
| A348ΔB1 to -B11 | A348 strains with nonpolar deletion of each of the 11 pTiA6NC virB genes | 7 |
| Plasmids | ||
| pUC118 | ColE1 cloning vector | Laboratory stock |
| pBSIIKS+ | ColE1 cloning vector | Laboratory stock |
| pBSKSgn | Gentamicin-resistant derivative of pBSIIKS+ | F. White |
| pAD1448 | phoA fusion vector; phoA codon 11 to end of gene in pUC118 | 15 |
| pGEX2T | ColE1 vector for GST fusion construction | GE Healthcare |
| pTJS75 | Wide-host-range IncP cloning vector | Laboratory stock |
| pAD1708 | pTJS75 derivative with the constitutive vir gene regulator virG(N54D) mutant | This study |
| pAD1412 | pTJS75Δ1 derivative with the virG(N54D) mutant | 15 |
| pAD1756 | virDp in pBSKSgn | This study |
| pAD1757 | virB3 as a SalI-XhoI fragment in pAD1756 | This study |
| pAD1758 | pAD1412 fused to pAD1757 | This study |
| pB3n-phoA | pAD1448 with a virB3-phoA fusion at the nth codon of virB3 | This study |
| pWB3n-phoA | pAD1412 fused to pB3n-phoA | This study |
| pGFP2 | virDp-gfp mut3 in pUC118 | This study |
| pGFP5 | pGFP2 with XbaI site filled in | This study |
| pGfp-virB3 | virB3 as an XbaI fragment in pGFP2 | This study |
| pVirB3-gfp | virB3 as an XhoI fragment in pGFP5 | This study |
| pAD1765 | pAD1708 fused to pGfp-virB3 | This study |
| pAD1784 | pAD1708 fused to pVirB3-gfp | This study |
| pGST-virB2 | virB2 codon 48 to end of gene in pGEX2T | This study |
| pAD1287 | pTiA6NC virB operon in pBSKSII+ | 2 |
| pTJ12 | virB1 to virB5 in pBSIIKS+ | This study |
| pAD1882 | virB1 to virB4 in pBSIIKS+ | This study |
| pAD1887 | pAD1412 fused to pAD1882 | This study |
| pAD1885 | virB1, virB2, and virB3-virB4 fusion in pBSIIKS+ | This study |
| pAD1888 | pAD1412 fused to pAD1885 | This study |
| pAD1889 | virBp-virB4 in pBSIIKS+ | This study |
| pAD1890 | pAD1412 fused to pAD1889 | This study |
| pAD1891 | virDp-virB2 in pGFP5 | This study |
| pAD1892 | pAD1412 fused to pAD1891 | This study |
| pAD1416 | virDp in pUC118 | This study |
| pAD1910 | virDp-virB7 in pAD1416 | This study |
| pAD1916 | pAD1412 fused to pAD1910 | This study |
| pAD1911 | virDp-virB8 in pAD1416 | This study |
| pAD1917 | pAD1412 fused to pAD1911 | This study |
| pAD1915 | virDp-virB7 and virB8 in pAD1416 | This study |
| pAD1918 | pAD1412 fused to pAD1915 | This study |
| pGEX2T | GST fusion vector | GE Healthcare |
| pGST-B2 | virB2 codon 48 to end of gene in pGEX2T | This study |