Figure 4
Analysis of transgene events. (A) DNA extracted from a set of T1 plants of each event subjected to DNA gel blotting, and probed with a DIG labeled fragment of HPT. Note that copy #3 of TG5E01 as revealed and indicated in Figure 3 was not highlighted by the HPT probe used here. (B) Detached leaf hygromycin assay 10 days after transfer to hygromycin-containing medium. (C) Amplification of the HPT coding region and the CaMV 35S terminator. (D) Amplification of the gfp coding region and the CaMV 35S terminator. (E) Duplex PCR targeting the CaMV 35S and the UBI-1 promoter, as well as the HPT and gfp coding sequence. (F) The T-DNA based transformation cassette. LB – left border, P35S – promoter of the CaMV 35S promoter, HPT – HYGROMYCIN PHOSPHOTRANSFERASE gene, T35S – terminator of the CaMV 35S coding sequence. TNOS – terminator of the NOPALINE SYNTHASE gene, gfp – synthetic green fluorescent protein gene (S65T), PUBI-int – maize UBI-1 promoter with first intron, RB – right border. The full length of the T-DNA, the relative position of the HindIII restriction site as well as the positions of probes used in A (HPT probe) and Figure 3 (gfp probe) as well as amplicons shown in C, D and E are indicated.