Dihydroflavonol-4-reductase (DFR) is a key enzyme of phenylpropanoid-flavonoid pathway. It plays important roles in pigmentation of seed coat and plant surface. A 600-bp (not including the artificial enzyme sites) RNAi fragment, BDFRI, targeted to Brassica DFR gene family was cloned from Brassica napus. Its antisense and sense fragments were inserted to the promoter-spacer and spacer-terminator cloning sites of plant RNAi platform vector pFGC5941 M which was modified by this research team recently, forming the 11400-bp RNAi vector pFGC5941 M-BDFRI. It was successfully verified by multiple PCR checking and then transformed into Agrobacterium tumefaciens strain LBA4404 to form engineering strain. Construction of this vector will lay a basis for clarifying the molecular mechanism of DFR in participating in the determination of Brassica seed coat pigments and stem/leaf colors and for metabolism regulation.%二氢黄酮醇-4-还原酶(dihydroflavonol-4-reductase,DFR)是类黄酮途径的一个关键酶,对种皮色泽和花色的形成具有重要作用.本研究从甘蓝型油菜克隆了600bp(不计人工酶切位点)的芸薹属DFR基因家族的RNA干扰片段BDFRI,将其反义片段和正义片段分别插入到本课题组新近改造的植物RNAi平台载体pFGGC5941M的启动子与间隔区之间、间隔区与终止子之间,构建了11400 bP的RNAi干扰载体pFGC5941M-BDFRI,通过多重PCR鉴定证实载体构建成功,并转化到根癌农杆菌菌株LBA4-404中形成了工程菌株.此载体的构建为进一步研究DFR基因参与决定芸薹属种皮色素和茎叶彩色性状的分子机理和代谢调控奠定了基础.
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