华北农学报Issue(4)：1-6,6.
大豆翻译起始因子4E 干扰片段克隆及其 RNAi载体构建
Cloning of Interference Fragment of Eukaryotic Translation Initiation Factor 4E of Soybean and Construction of Its RNAi Vector
摘要
Abstract
Soybean transformation based on RNA interference ( RNAi) can specifically silence the homologous target genes in mRNA level ,providing a new strategy for disease-resistant breeding .In this study ,we made the ami-no acid sequence alignment of eIF4E from ten different kinds of plants which have been identified to interact with the virus VPg ,then we determined the conserved interval of soybean eIF 4 E was 62-237 amino acid sequence and cloned the 406 bp interference fragment named eIF4Ei (including 58 bp specific recombination sequence attB ).U-sing the GATEWAY technology ,the RNAi vector pB7GWIWG2(Ⅱ)-eIF4Ei was constructed,which was identified by PCR amplification ,sequencing and restriction digestion .The identification of the recombinant expression vector ensured the eIF4 Ei interference fragment no mutation in the process of vector construction and two ccdB sites were all replaced by eIF4Ei,in addition,the insert direction of the two interference fragments in the terminal plasmid was conversed.Overall,the RNAi vector was successfully constructed to be an inverted repeat structure ,providing the basic materials for cultivating new germplasm of soybeans resistant to SMV using the soybean transformation technol -ogy based on RNAi .关键词
大豆花叶病毒/eIF4E基因/RNA干扰/GATEWAY技术/载体构建Key words
Soybean mosaic virus/eIF4E gene/RNA interference/GATEWAY technology/Vector construction分类
农业科技引用本文复制引用
高乐,翟锐,丁雪妮,李凯,章红运,王涛,智海剑..大豆翻译起始因子4E 干扰片段克隆及其 RNAi载体构建[J].华北农学报,2014,(4):1-6,6.基金项目
转基因生物新品种培育科技重大专项（2008ZX08004-004）；国家自然科学基金项目（31171574；31371646；31101164）；农业部大豆产业技术体系项目 （）
