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RNAi系双链RNA诱导同源mRNA降解,导致特定基因表达沉默的一种现象.RNAi技术是通过基因工程防治植物病毒病的最佳途径.由大麦黄矮病毒(Barely yellow dwarf virus,简称BYDV)引起的禾谷类黄矮病发病面积和危害程度在我国呈加重趋势.BYDV-GAV是当前流行的大麦黄矮病毒株系,本文针对BYDV-GAV复制酶基因序列,构建能在细胞内转录形成发卡RNA双链结构的表达盒,并将其置于中间载体pVec8-2b的T-DNA区,pVec8-2b的另一T-DNA区含有hpt选择报告基因表达盒.具双T-DNA区的病毒复制酶基因发卡RNA高效表达载体已导入根癌农杆菌,可用于诱导植物RNAi,创制无选择标记基因的抗大麦黄矮病转基因作物.图6参21
RNAi double-stranded RNA induces the degradation of homologous mRNA, resulting in the silencing of specific gene expression.RNAi technology is the best way to prevent and cure plant virus disease by genetic engineering.By the method of Bark disease of yellow dwarf virus (BYDV) The incidence and harm degree of cereal yellowish dwarf disease in our country is aggravating trend.BYDV-GAV is the prevailing barley yellow dwarf virus strain. In this paper, BYDV-GAV replicase gene sequence, the construction can be transcribed in the cell to form a card RNA double-stranded construct and placed in the T-DNA region of the intermediate vector pVec8-2b containing the hpt selection reporter cassette in the other T-DNA region. Replicase gene hairpin RNA efficient expression vector has been introduced into Agrobacterium tumefaciens, can be used to induce plant RNAi, the creation of non-selectable marker gene anti-barley yellow dwarf transgenic crops.