| TMV和PVY外壳蛋白双基因融合干涉及烟草转化 |
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| 引用本文： | 刘丹,郭兆奎,万秀清,颜培强,乔婵,刘磊.TMV和PVY外壳蛋白双基因融合干涉及烟草转化[J].广西农业生物科学,2009(3):450-454. |
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| 作者姓名： | 刘丹 郭兆奎 万秀清 颜培强 乔婵 刘磊 |
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| 作者单位： | 黑龙江八一农垦大学植物科技学院;牡丹江省烟草科学研究所; |
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| 基金项目： | 国家烟草专卖局科技计划项目资助 |
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| 摘 要： | 为了降低烟草花叶病毒（tobacco mosaic virus，TMV）和马铃薯Y病毒（potato irus Y，PVY）复合侵染对烟草带来的危害，本实验找到TMV—CP和PVY—CP基因部分保守序列，将保守序列进行双基因融合，此双基因即为RNAi的靶序列，用限制性内切酶将双基因从pMD18-T载体上切下，正反向连接到pUCCRNAi载体后，经酶切鉴定后定向连接到含超强启动子的pC2300—35S—OCS表达载体上，利用冻融法将此表达载体导入只含辅助质粒的根癌农杆菌中，构建含靶序列反向重复结构的RNAi双元载体系统，提取转化质粒，经酶切验证鉴定表明TMV和PVY外壳蛋白基因植物表达双元载体构建成功。并转化烟草，获得了3株对TMV和PVY抗性显著提高的转基因烟草。
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| 关 键 词： | TMV和PVY双基因融合 RNA干涉 转化烟草 |
RNA Interference of TMV-CP and PVY-CP Double Gene to Fusion and Transform Tobacco |
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| Liu Dan Guo Zhaokui , Wan Xiuqing Yan Peiqiang Qiao Chan Liu Lei Heilongjiang August First L, Reclamation University,Daqing, Mudanjiang Tobacco Research Institute,Mudanjiang.RNA Interference of TMV-CP and PVY-CP Double Gene to Fusion and Transform Tobacco[J].Journal of Guangxi Agricultural and Biological Science,2009(3):450-454. |
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| Authors: | Liu Dan Guo Zhaokui Wan Xiuqing Yan Peiqiang Qiao Chan Liu Lei Heilongjiang August First L Reclamation University Daqing Mudanjiang Tobacco Research Institute Mudanjiang |
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| Institution: | Liu Dan 1 Guo Zhaokui 1,2 Wan Xiuqing 2 Yan Peiqiang 2 Qiao Chan 2 Liu Lei 1 1 Heilongjiang August First L, Reclamation University,Daqing,163319,2 Mudanjiang Tobacco Research Institute,Mudanjiang,157011 |
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| Abstract: | The objective is to decrease the dangerous that the TMV(tobacco mosaic virus) and PVY(potato virus Y) virus complex infect tobacco.Firstly we found the conserved sequence of TMV-CP gene and PVY-CP gene respectively.Then they were cloned and fusioned in to pMD18-T vector as the target gene.Secondly we construct RNA interference expression plasmid of target gene by cloning reverse repeats into plasmid DNA and then plant expression vector pC2300-35s-OCS was linked with it.The plant expression vector was transf... |
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| Keywords: | TMV-CP and PVY-CP gene fusion RNA interference Transforme tobacco |
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