Figure 4
Molecular characterizations of Bt-transgenic progenies and receptor binding assay. A RT-PCR analysis of 10 selected transgenic tomato plants from T2 to T6 generation of T1 transgenic plant Ab25.3. M: 1kb ladder, cry1Ab amplicon size is 800 bp. B Southern blot hybridization of Eco RI digested DNA from transgenic tomato Ab25 E line, in T4, T5 and T6 generation, +C 1845 bp fragment of cry1Ab gene as positive control, –C: Genomic DNA from non-transformed tomato as negative control. C Western blot with protein extract from Ab25 E transgenic tomato line expressing cry1Ab gene in T4, T5 and T6 generations, +C: partially purified Bt-protein of ~65 kDa, –C as positive control: Total protein isolated from non-transformed tomato plant as negative control. D Binding assay of plant expressed Cry1Ab toxin protein with BBMV protein of H. armigera (■) and S. litura (○). The BBMV protein fraction (300 ng/100 μl) from both the insects were coated and shown in long scale while Cry1Ab toxin protein (125 ng/ 100 μl) was used for protein-protein interaction followed by immuno absorbance assay.