Figure 8

Schematic structure of T-DNA region of overexpression vector used for E. ulmoides transformation. cDNA of EuIPI was inserted into pMSIsGFP binary vector between 35S promoter and a NOS terminator. NPT II gene was used as a selective marker, and sGFP(S65T) gene was used to optimize conditions for transformation by monitoring the expression of green-fluorescent protein. An intron was fused within the N-terminal part of the sGFP(S65T) coding sequence to discriminate between Agrobacterium and plant expression (because bacteria can not splice the intron). RB, right border; LB, left border; NOS-P, nopaline synthase promoter; NOS-T, nopaline synthase terminator; 35S-P, cauliflower mosaic virus (CaMV) 35S promoter; 35S-Ω-P, 35S promoter with additional omega element translational enhancer; NPT II, neomycin phosphotransferase; sGFP(65T), synthetic green-fluorescent protein with S65T mutation; I: intron of castor bean catalase gene CAT-1.