含融合杀虫基因和VHb基因双价表达载体的构建  

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

作　　者：王友如[1,2] 郭三堆[2] 

机构地区：[1]湖北师范学院生命科学学院,湖北黄石435002 [2]中国农业科学院生物技术研究所基因工程实验室,北京100081

出　　处：《生物技术》2010年第3期6-8,共3页Biotechnology

摘　　要：目的:探索分子育种的新途径,获得高产多抗虫谱的转基因植物。方法:将密码子优化后的透明颤菌血红蛋白(vgbM)基因与融合杀虫基因(GFMcryIA+CPTI)构建成双价基因植物高效表达载体pGBI4ABCVHB,基因表达盒中含2个增强子、35S启动子、Ω前导序列、Kozak序列、多联终止密码子及Nos终止子以及正确切割、加工序列、Poly(A)信号序列。结果:通过根癌农杆菌介导转化烟草,获得了转基因植株,结论:vgbM基因在转基因烟草中有效表达;融合杀虫基因也表达出活性毒蛋白。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.

关 键 词：融合杀虫基因 VHB基因 双价表达载体 抗虫性 活性蛋白 

分 类 号：Q782[生物学—分子生物学]