Construction of Bivalent Gene Expression Vector Harbouring VHb and Fused Insectidal Gene

Objective:To obtain transgenic plan with high-yiled and multiple spectrum of insect-resistance by molecular breeding technology,the highly-efficient plant expression vector harbouring bivalent genes was constructed by applying the artificially-modified-synthesized VHb gene(vgbM gene) and the fused-insectidal gene(GFMcryIA + CPTI),vgbM gene experession cassettes contained such cis-acting elements as 35S promoter,two enhancers,Ω and Kozak sequence,poly-terminator and Nos terminator,besides these elements,the fused-insectidal gene expression cassettes contained splicing sequence,processing sequence and Poly(A) sequence.Result:Transgenic tobacco plants were obtained through the transformation with Agrobacterium tumefaiens carrying the expression vector pGBI4ABCVHB,by PCR amplification and Southern blot analysis,it was proved that the insectidal genes were inserted into the tobacco plant genome;through western blot analysis it was also verified that vgbM gene expressed in the transgenic plants.Conclusion:Toxicity assay indicated that transgenic plants expressed pesticidal toxin protein.