高效Bt抗虫基因表达结构的构建及其在转基因烟草中表达行为的研究  

Construction of Plant Vectors with High Level Expression of Bt Toxin Gene and Studies on Their Expressions Behavior in Transgenic TobaccosSupported by the High Technology Research and Development Programme of China and the National Natural Scien

作　　者：石春林[1] 朱祯[1] 肖桂芳[1] IllimarAltosaar 冯平章[3] 

机构地区：[1]中国科学院遗传研究所,北京100101 [2]DepartmentofBiochemistry,OttwaUniversity [3]中国农业科学院植物保护研究所

出　　处：《生物工程学报》1999年第4期422-427,共6页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划项目;国家自然科学基金

摘　　要：构建了高效植物表达载体ｐ Ｂｉｎ Ｍｏ Ｂｃ ，其携带有超强表达复合启动子 Ｏ Ｍ 及Ω因子控制下的 Ｃｒｙ Ｉ Ａ（ｃ） 基因，作为对照，本实验构建了含有 Ｃａ Ｍ Ｖ３５ Ｓ 启动子控制下的 Ｃｒｙ Ｉ Ａ（ｃ）基因的植物表达载体ｐ Ｂｉｎｏ Ｂｃ 。分别使用两个植物表达载体转化烟草， Ｅ Ｌ Ｉ Ｓ Ａ 检测表明，在ｐ Ｂｉｎ Ｍｏ Ｂｃ 转基因烟草中 Ｃｒｙ Ｉ Ａ（ｃ） 基因的平均表达水平是ｐ Ｂｉｎｏ Ｂｃ 的２４４ 倍，最高可达可溶蛋白的０２５５ ％ 。抗虫检测结果表明，ｐ Ｂｉｎ Ｍｏ Ｂｃ 转基因烟草与ｐ Ｂｉｎｏ Ｂｃ 转基因烟草相比，具有更强的抗棉铃虫效果。上述结果表明， Ｏ Ｍ 启动子比 Ｃａ Ｍ Ｖ ３５ Ｓ 启动子更具有实际应用价值，此结果在植物抗虫基因工程研究中具有重要意义。Breeding pest resistant plants using plant genetic engineering technique is an effective strategy in the integrated pest management(IPM).Increasing the expression level of foreign insecticidal protein by using strong promoter is a useful method.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.An another vector,pBinoBc,was also constructed in this study.pBinoBc caring Cry IA(c) gene was under control of CaMV 35S promoter.The vectors were transformed into toba cco respectively.ELISA assay showed that the expression level of the Cry IA(c) gene in pBinMoBc transgenic tobacco plants is 2.44 times that in pBinMoBc transgenic tobacco plants,the expression level of Bt toxin can be up to 0.255% of total soluble proteins.Bioassay showed that pBinMoBc transgenic tobacco plants had more notable insecticidal effects than pBinMoBc transgenic tobacco plants.The above result showed that the chimeric OM promoter is a stronger promoter than CaMV 35S promoter that was widely used in plant genetic engineering,and this is very useful in pest resistant plant genetic engineering.

关 键 词：转基因植物 抗虫基因 复合OM启动子 抗虫检测 

分 类 号：Q785[生物学—分子生物学] S572.034[农业科学—烟草工业]