The BmK IT4 gene was obtained from pBS-BmK IT4 by EcoRⅠ/KpnⅠdigestion and it was then cloned into pE3 intermediate vector. The resulting plasmid was named pE3-BmK IT4. The chimeric gene was transferred into the tobacco (Nicotiana tabacum L.) genome via Agrobacterium-mediated transformation. Forty-five regenerated kanamycin resistant plants were obtained, two individual lines showed strong toxicity to Manduca sexta (Linnaeus), Heliothis armigera (Hübner) and Leguminivora glycinivorella (Matsumura) by feeding experiments. Results from Southern blot indicated that BmK IT4 gene was transferred into tobacco genome. The mortality of M．sexta, H．armigera and L．glycinivorella larvae fed on transgenic plants was 95%-97%, 63%-70% and 65%-73%, respectively, and the growth of the surviving insects was remarkably retarded.
转BmK IT4基因烟草的抗虫性
胡廷章1,2 徐云剑1 江国英1 朱玉贤1*
（1. 北京大学生命科学学院蛋白质工程与植物基因工程国家重点实验室，北京10081；
2. 四川三峡学院生物系，重庆404000）
摘要：用酶切方法从pBS-BmKIT4 质粒中获得BmKIT4 基因片段 ,并构建CaMV 35S启动子下的BmKIT4 基因表达质粒pE3_BmKIT4 ,以根癌土壤杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)介导的叶盘法转化云烟 (Nico tianatabacumL .)K32 6叶片 ,获得 45株抗卡那霉素的再生植株。用这些再生植株进行抗虫试验 ,发现有 2株植株对烟草天蛾 (Manducasexta (Linnaeus) )、棉铃虫 (Heliothisarmigera (H櫣ｂｎｅｒ) )和大豆食心虫 (Leguminivoraglycinivorella(Matsumura) )都有较强的抗性。收获这 2株烟草的自交种子后 ,对T1代幼苗进行Southern杂交分析表明 ,BmKIT4基因已转入这些烟草中 ,用这些子代植株进行抗虫试验 ,与对照株相比 ,这 2株转基因烟草对烟草天蛾的杀虫率达95 %～ 97% ,对棉铃虫的杀虫率达 6 3%～ 70 % ,对大豆食心虫的杀虫率达 6 5 %～ 73% ,并能显著抑制昆虫的蜕皮和生长发育 ,表现出强烈的抗虫作用。
关键词： BmKIT4 基因；转基因烟草；抗虫性
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