优化的VHb基因和融合杀虫基因在烟草中的表达.pdfVIP

农业生物技术学报 Journal ofAgricultural Biotechnology 2007，1 5 (6)：982～986 · 研究论文· 优化的VHb基因和融合杀虫基因在烟草中的表达水 王友如-， ¨，崔洪志2，郭三堆2 (1．湖北师范学院，黄石435002；2．中国农业科学院生物技术研究所，北京100081) 摘要：将人工合成的、密码子优化后的透明颤菌血红蛋白(VI4b)基因(vgbM)与含有融合杀虫基因(GFMcrylA和cP订)表 达载体pGBIF4ABC构建成双价基因植物高效表达载体pGBIF4ABCVHB，通过根癌农杆菌(Agrobacterium tumefaciens)介导转 化烟草(Nicotiana tabacmn)，经PCR及Southern blot检测，证实了双价基因在32株转基因烟草基因组中整合。We~em blot检测 证实了vgbM基因的表达，杀虫实验表明，融合杀虫基因表达的毒蛋白具有杀虫活性，转基因烟草平均每株干重高于非转基因 烟草植株8％。表明VHb基因和融合杀虫基因烟草在烟草中的表达可使烟草可增产。又具抗虫性。 关键词：V／-／b基因；融合杀虫基因；转基因植株；抗虫性；产量 中图分类号：SI88文献标识码：A 文章编号：1006．1304(2007)06．0982-05 Expression of Modified VHb Gene and Fusedly-insectidal Genes in Tobacco WANY You-ru ， ¨，CUI Hong-zhi ，GUO San-dui HubeiNormal College,H~mgshi 435002,China； ． 2．BiotechnologyResearchInstitute,Ch／neseAcademyofAgricultmalSciences,Beifing100081，ChinaJ Abllllll~：The highly-efficient plant binary expression vector pGBIF4ABCVHB was constructed by applying the artificially-modified-synthesized vgbM gene and fusedly-insectidal genes(GFMcry／A and CPT／)．Transgenic tobacco(Nicotiana tabacmn)plants were obtained by Agrobacterium mediated transformation，32 ofthem were identified that the bivalent genes were inse~ed into the tobacco genome by PCR amplification deletion and Southern blotting analysis．Western blotting analysis proved that vgbM gene Was expressed in the transgenic plants．Toxicity assay indicated that fused insectidal genes were expressed activity of pesticadal toxin protein．The net weight of transgenic tobacco plants exceeded 8％ of that of non-transgenic ones．Compared to non- transgenic tobacco plants，the transgenic ones were high-yield varieties ofinsect-resistance． K 神 ：V／-／b gene；fusedly-insectidal ge