| 油菜叶绿体定点转化载体的构建及其杀虫性 |
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| 引用本文： | 侯丙凯,张中林,周奕华,章银梅,沈桂芳,陈正华. 油菜叶绿体定点转化载体的构建及其杀虫性[J]. 高技术通讯, 2000, 10(7): 5-11 |
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| 作者姓名： | 侯丙凯 张中林 周奕华 章银梅 沈桂芳 陈正华 |
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| 作者单位： | 1. 中国科学院遗传研究所,北京,100101
2. 中国农业科学院生物技术研究所,北京,100081 |
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| 基金项目： | 国家自然科学基金!(39970624) |
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| 摘 要： | 首先从油菜叶绿体基因组克隆得到包含ｒｐｓ７基因在内的１．０ｋｂＤＮＡ片 包含ｎｄｈＢ基因在内的２．４ｋｂＤＮＡ片段，同时从苏云芽孢杆菌质粒上克隆得到一个全长３．５ｋｂ的ＢＴ杀虫蛋白基因ｃｒｙ１Ａａ。然后以ｒｐｓ７和ｎｄｈＢ基因作为同源重组片段，成功构建了包含Ｂｔ杀虫蛋白和ａａｄＡ抗壮观霉素基因的油菜叶绿体定点转化载体，并对克隆菌体总蛋白进行了生物杀虫试验。结果表明，Ｂｔ杀虫蛋白基因能够得到表达，并
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| 关 键 词： | 油菜 叶绿体转化 载体构建 杀虫试验 |
Construction of Vector for Oilseed Rape Chloroplast Transformation and Its Insecticidal Toxicity |
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| Hou Bingkai,Zhang Zhonglin,Zhou Yihua,Zhang Yinmei,Shen Guifang,Chen Zhenghua. Construction of Vector for Oilseed Rape Chloroplast Transformation and Its Insecticidal Toxicity[J]. High Technology Letters, 2000, 10(7): 5-11 |
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| Authors: | Hou Bingkai Zhang Zhonglin Zhou Yihua Zhang Yinmei Shen Guifang Chen Zhenghua |
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| Abstract: | WT5BZ]1.0 kb and 2.4 kb DNA fragment containing rps7 gene and ndhB gene, respectively, were cloned from oilseed rape chloroplast genome. A full length 3.5 kb Bt toxin gene ( cry1Aa ) was also cloned from the plasmid of Bacillus thuringiensis. The vector containing Bt toxin gene and aadA gene for oilseed rape chloroplast transformation was constructed with the rps7 gene and ndhB gene as homologous recombinant fragments. Bioassays using crude expressed products from host strain of the vector showed that Bt toxin gene was expressed and its products were toxic to larvae of Heliothis armigera. The chloroplast transformation vector will be of great value to oilseed rape chloroplast transformation and to the insect resistant crop improvement. |
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| Keywords: | Oilseed rape Chloroplast transformation Vector construction Insecticidal test |
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