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当前位置: 首页 > 文章 > 靶向CMV外壳蛋白基因的RNA干涉在番茄中转化及表达 北方园艺 2011 (6) 133-136
Position: Home > Articles > Transform and Expression of RNA Interferential Targeting CMV-Coat Protein Gene in Tomato Northern Horticulture 2011 (6) 133-136

靶向CMV外壳蛋白基因的RNA干涉在番茄中转化及表达

作  者：
潘永明;张李华;李春彦;周岩
单  位：
牡丹江医学院;牡丹江师范学院;密山市8511农场林业局
关键词：
RNA干涉;黄瓜花叶病毒;基因转化;基因表达
摘  要：
以CMV(黄瓜花叶病毒)外壳蛋白基因为靶标基因,针对CMV 3个常见株系(普通株系、黄化株系和坏死株系)外壳蛋白基因间高度同源序列设计引物,经RT-PCR获得目的序列,构建RNA干涉双元载体(其中目的序列反向重复插入)。以含双元载体的根癌农杆菌介导转化无菌番茄苗,培育转基因番茄。人工接种CMV验证转基因番茄抗病毒能力,荧光定量PCR分析转基因番茄中外源基因的表达效果。结果表明:RNA干涉转基因番茄呈现不同程度的抗病毒能力;对表现抗病毒能力的转基因番茄进行荧光定量PCR分析显示,转基因番茄中靶标基因的转录mRNA存在一定程度的降解,且抗病毒能力与降解程度成正相关。
译  名：
Transform and Expression of RNA Interferential Targeting CMV-Coat Protein Gene in Tomato
作  者：
PAN Yong-ming1,ZHANG Li-hua2,LI Chun-yan3,ZHOU Yan1(1.Mudanjiang Normal University,Mudanjiang,Heilongjiang 157012;2.Mishan 8511 Farm Forestry Bureau,Mishan,Heilongjiang 158308;3.Mudanjiang Medical University,Mudanjiang,Heilongjiang 157011)
关键词：
RNA interference;cucumber mosaic virus;gene transformation;gene expression
摘  要：
The coat protein gene of cucumber mosaic virus(CMV) was targeted for RNA interference in this paper.The partial sequence of CMV coat protein gene cDNA was obtained by the RT-PCR with the primers which designed according to the high homologous sequences of three CMV three strains.The target sequences were prepared by PCR and inverted repeat in plasmid to build the RNA interference binary vector.The tomato were genetic modified via binary vector of Agrobacterium tumefaciens-mediated transformation.The resistance to CMV of transgenic tomato was identified by artificial inoculation.The expression of RNA interference effect of the transgenic tomato were evaluated by realtime PCR analysis.The results showed that RNAi-transgenic tomato showed ability of different levels of anti-virus,the quantitative PCR analysis for transgenic tomatoes antiviral capacity showed that the transgenic tomatoes target gene transcription mRNA exist a certain degree of degradation,and the degree of degradation was positive correlation to antiviral capacity.

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