| Abstract: | By in situ hybridization of bacterium clone and analysis of restriction enzyme digestion, both CMV-cp gene and Bt-toxin gene were inserted one by one into T-DNA of binary plant expression vector pea. The reconstructed plasmid was named pE14. Then, tomato was transformed with pE14 mediated by Agrobacterium tumefaciens GV311-SE, four regenerated tomato plants were obtained on the MS medium containing 100 μg/mL kanamycin. Assay of nopaline, dot blotting of tomato genomic DNA and PCR amplication of CMV-cp gene and Bt-toxin gene from genomic DNA showed that CMV-cp gene and Bt-toxin gene were transferred into the four regenerated tomato plants simultaneously with T-DNA, and no recombination of genes occurred. RNA dot blotting showed that two of them could express simultaneously the CMV-cp gene and Bt-toxin gene proteins. The resistances to virus and insect of the transgenic tomato plants will be tested in their F1 and F2 regenerations. |
