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Cloning of Cotton Delta-12 Oleate Desaturase Gene FAD2-1 and Construction of Its ihpRNA and amiRNA Interference Vectors 被引量:1

棉花FAD2-1基因的克隆及其ihpRNA和amiRNA干扰载体的构建(英文)
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摘要 Delta-12 oleate desaturase gene (FAD2-1) which converts oleic acid into linoleic acid, is the key enzyme determining the fatty acid composition of cottonseed oil. By employing RT-PCR method, full length cDNA of cotton delta-12 oleate desat- urase gene GhFAD2-1 containing an open reading frame of 1 158 bp was cloned for constructing RNAi vector. A 515 bp long specific fragment of this gene was se- lected for constructing ihpRNA vector under the control of a seed-specific promoter NAPIN, named pFGC1008-NAPIN-FAD2-1; meanwhile miRNA gene-silencing vector pCAMBIA1302-amiRNA-FAD2-1 targeting GhFAD2-1 was also constructed. 采用RT-PCR方法克隆了棉花-12脂肪酸去饱和酶基因(GhFAD2-1)的cDNA全长序列,该基因含有1158bp的完整开放阅读框。棉花FAD2-1是种子中编码催化油酸形成不饱和脂肪酸的关键酶的基因,选取GhFAD2-1基因中的一段特异的515bp片段,构建了种子特异性启动子NAPIN调控的ihpRNA干扰表达载体,pFGC1008-NAPIN-FAD2-1,同时构建了针对GhFAD2-1基因的人工miRNA表达载体pCAMBIA1302-amiRNA-FAD2-1。
出处 《Agricultural Science & Technology》 CAS 2012年第11期2281-2283,2286,共4页 农业科学与技术(英文版)
关键词 Cotton delta-12 oleate desaturase gene GhFAD2-1 ihpRNA interferencevector amiRNA interference vector High oleic acid contents 脱氢酶基因 亚油酸 RNA干涉 载体 棉花 克隆 ami RT-PCR方法
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参考文献9

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