TABLE 1.

Bacterial strains and plasmids used for this study

Strain or plasmidRelevant characteristicsSource or reference
Strains
    E. coli
        DH5αF′F′ endA1 hsdR17 recA1 gyrA Δ(lacZYA-argF)U169 φ80dlacZΔM15Laboratory stock
        CJ236FΔ(HindIII)::cat ung-1 dut-1Laboratory stock
    Agrobacterium tumefaciens
        C58Harbors nopaline Ti plasmid pTiC58Laboratory stock
        A136C58 cured of pTiC58Laboratory stock
        A348A136 with octopine Ti plasmid pTiA6NCLaboratory stock
        A281A136 with succinamopine Ti plasmid pTiBo542Laboratory stock
        AD802phoA derivative of A13615
        A348ΔBA348 with a deletion of pTiA6NC virB operon (PC1001 to PC1011)2
        A348ΔB1 to -B11A348 strains with nonpolar deletion of each of the 11 pTiA6NC virB genes7
Plasmids
    pUC118ColE1 cloning vectorLaboratory stock
    pBSIIKS+ColE1 cloning vectorLaboratory stock
    pBSKSgnGentamicin-resistant derivative of pBSIIKS+F. White
    pAD1448phoA fusion vector; phoA codon 11 to end of gene in pUC11815
    pGEX2TColE1 vector for GST fusion constructionGE Healthcare
    pTJS75Wide-host-range IncP cloning vectorLaboratory stock
    pAD1708pTJS75 derivative with the constitutive vir gene regulator virG(N54D) mutantThis study
    pAD1412pTJS75Δ1 derivative with the virG(N54D) mutant15
    pAD1756virDp in pBSKSgnThis study
    pAD1757virB3 as a SalI-XhoI fragment in pAD1756This study
    pAD1758pAD1412 fused to pAD1757This study
    pB3n-phoApAD1448 with a virB3-phoA fusion at the nth codon of virB3This study
    pWB3n-phoApAD1412 fused to pB3n-phoAThis study
    pGFP2virDp-gfp mut3 in pUC118This study
    pGFP5pGFP2 with XbaI site filled inThis study
    pGfp-virB3virB3 as an XbaI fragment in pGFP2This study
    pVirB3-gfpvirB3 as an XhoI fragment in pGFP5This study
    pAD1765pAD1708 fused to pGfp-virB3This study
    pAD1784pAD1708 fused to pVirB3-gfpThis study
    pGST-virB2virB2 codon 48 to end of gene in pGEX2TThis study
    pAD1287pTiA6NC virB operon in pBSKSII+2
    pTJ12virB1 to virB5 in pBSIIKS+This study
    pAD1882virB1 to virB4 in pBSIIKS+This study
    pAD1887pAD1412 fused to pAD1882This study
    pAD1885virB1, virB2, and virB3-virB4 fusion in pBSIIKS+This study
    pAD1888pAD1412 fused to pAD1885This study
    pAD1889virBp-virB4 in pBSIIKS+This study
    pAD1890pAD1412 fused to pAD1889This study
    pAD1891virDp-virB2 in pGFP5This study
    pAD1892pAD1412 fused to pAD1891This study
    pAD1416virDp in pUC118This study
    pAD1910virDp-virB7 in pAD1416This study
    pAD1916pAD1412 fused to pAD1910This study
    pAD1911virDp-virB8 in pAD1416This study
    pAD1917pAD1412 fused to pAD1911This study
    pAD1915virDp-virB7 and virB8 in pAD1416This study
    pAD1918pAD1412 fused to pAD1915This study
    pGEX2TGST fusion vectorGE Healthcare
    pGST-B2virB2 codon 48 to end of gene in pGEX2TThis study