Figure 1
Schematic diagram of the T-DNA region of binary vector pBIN200 and molecular characterization of T 0 and T 1 transgenic tomato plants. A T-DNA region of pBIN200 harbouring cry1Ab gene driven by constitutive DECaMV35S promoter. Different PCR primers used in the present study are shown with arrow marks at their respective binding positions: npt II-F, npt II-R for npt II gene and Ab-F, Ab-R for cry1Ab gene amplification, respectively. Bold line represents 1845 bp Bam HI and Eco RI fragment of cry1Ab gene used for preparation of radiolabelled probe for Southern blot hybridization analysis in the present study. RB and LB: right and left border sequences, npt II: neomycin phosphotransferase gene, AMV: alfalfa mosaic virus 5′ UTR sequence, cry1Ab: modified truncated 1845 bp cry1Ab gene of B. thuringiensis, DECaMV35S: CaMV35S promoter with duplicated enhancer, Pnos: nos promoter, Tnos: nos terminator. PCR amplification of genomic DNA from transgenic tomato in T0 and T1 generations. B, F 678 bp amplification of npt II gene. C, G 800 bp amplification of cry1Ab gene. D, H RT-PCR analysis of T0 and T1 transgenic tomato showing 800 bp amplicon of cry1Ab gene transcripts, respectively, +C: truncated 1845 bp cry1Ab gene as positive control. –C: DNA from non-transformed plant as negative control. E, I Southern blot analysis of PCR positive T0 transgenic plants and randomly selected T1 transgenic plants. +C: 1845 bp fragment of truncated cry1Ab gene. –C: DNA from non-transformed control plant.