Bt toxin genes were the insecticidal genes most widely used in genetic engineering of pest resistant plant, were of important significance to study their expression behavior in transgenic plants. In this work, a plant expression vector, pBinMoBc, was constructed. It contained the Cry IA(c) gene under control of chimeric OM promoter and the Ω factor. The vector was transferred into tobacco (Nicotiana tabacum L.) plant via Agrobacterium-mediated transformation. ELISA assay showed that the expression levels of the Cry IA(c) gene in transgenic tobacco plants were significantly higher than that in wild-type tobacco plants. The highest could be up to 0.255% of total soluble proteins; the expression level of CryIA(c) gene in transgenic tobacco plant was changeable during the development stages of tobacco plant. Bioassay showed that pBinMoBc transgenic tobacco plants had more notable insecticidal activity than the wild-type tobacco plants. The above results indicated that pBinMoBc was an effective pest-resistent plant expression vector. This study would be very helpful in screening transgenic cotton with high resistance to cotton bollworm (Heliothis armigeva Hubner).
转基因烟草中Bt 毒蛋白基因的表达行为
石春林 朱祯* 徐鸿林 肖桂芳 冯平章**
（中国科学院遗传研究所，北京100101）
摘要：构建了高效植物表达载体pBinMoBc ,该载体携带超强表达复合启动子OM及Ω因子控制下的CryIA(c)基因。采用根癌土壤杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)介导的方法转化烟草 (Nicotianatabacum L .) ,ELISA检测表明 ,大多数转基因烟草中CryIA(c)基因表达量均超过 0 .1% ,最高可达 0 .2 5 5 % ;转基因烟草中CryIA(c)基因表达水平与植株发育的时间和空间有关。抗虫检测结果表明 ,转基因烟草具有很强的抗棉铃虫 (He liothisarmigevaHubner)效果。上述结果表明 ,pBinMoBc是一个高效抗虫植物表达载体。
关键词： 转基因植物；Bt 毒蛋白基因；高效启动子；ELISA检测；抗虫检测
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