| 外源RNA干涉基因在烟草中的转化及表达 |
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| 引用本文： | 律凤霞,潘永明,郭兆奎.外源RNA干涉基因在烟草中的转化及表达[J].西北植物学报,2009,29(10). |
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| 作者姓名： | 律凤霞 潘永明 郭兆奎 |
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| 作者单位： | 律凤霞,潘永明(牡丹江师范学院,生物系,黑龙江牡丹江,157012);郭兆奎(黑龙江省烟草科技研究所,黑龙江牡丹江,157011) |
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| 基金项目： | 国家烟草专卖局科技项目 |
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| 摘 要： | 依据RNA干涉机制,以TMV复制酶基因为靶标基因,针对TMV 5个株系复制酶基因间高度同源序列设计引物,经RT-PCR反应获得靶序列,构建靶序列反向重复结构的RNA干涉双元载体.用根癌农杆菌介导将外源基因转化至烟草品种K326基因组中,培育RNA干涉转基因烟草.人工接种病毒验证转基因烟草中外源基因在植物抗病毒能力方面的表达效果,实时荧光定量PCR分析转基因烟草抗病毒能力.结果表明,实验培育的RNA干涉转基因烟草67%对TMV呈现高度抗性;荧光定量PCR分析显示,对TMV具高度抗性的转基因烟草中病毒复制酶基因转录产物mRNA存在很大程度的降解,证实了RNA干涉技术在培育抗病毒烟草品种中的效果.
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| 关 键 词： | RNA干涉 烟草花叶病毒 载体构建 基因表达 |
Transform and Expression of Exogenous RNA Interferential Gene in Tobacoo |
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| Abstract: | The aim of this paper is to investigate the transformation and expression of RNA interference in tobacco.TMV RNA was isolated and TMV-replicase cDNA sequence was obtained by the RT-PCR.The high homologous sequences were obtained by multiple alignmented of five TMV strains.The target sequences were prepared by PCR reaction with the primers which was designed according to the high homologous sequences.And the inverted repeat target sequence was used to build the RNA interference binary vector.The tobacco vari... |
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| Keywords: | RNA interference tobacco mosaic virus vector construction gene expession |
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