具双T-DNA区的大麦黄矮病毒复制酶基因RNAi植物表达载体的构建(英文)  

Construction of Twin T-DNA Vector Encoding Hairpin RNA Targeting the Replicase Gene of Barley Yellow Dwarf Virus

作　　者：孙文瑜[1,2] 陈静[3] 王磊[4] 付体华[2] 

机构地区：[1]中国科学院成都生物研究所成都610041 [2]四川农业大学农学院 [3]中国科学院成都生物研究所 [4]中国农业科学院生物技术研究所

出　　处：《应用与环境生物学报》2008年第6期745-749,共5页Chinese Journal of Applied and Environmental Biology

基　　金：Supported by the National Natural Science Foundation of China (No. 30871527);the Knowledge Innovation Project of the Chinese Academy of Science

摘　　要：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.RNA interference （RNAi） technology has become the most effective approach for genetically engineering virus resistance in plants. Here, we constructed a hairpin RNA （hpRNA）-expressing RNAi vector targeting the replicase gene of a Chinese isolate of barley yellow dwarf virus （BYDV）, BYDV-GAV. This construct was comprised of two tandem T-DNAs （twin T-DNAs）, one of which contained the hygromycin resistance gene hpt as the selectable marker, and the other contained the expression cassette encoding hpRNA of the polymerase gene sequence of BYDV-GAV. This twin T-DNA construct should allow the production of selectable marker-free transgenic plants that are resistant to BYDV-GAV. Fig 6, Ref 21

关 键 词：大麦黄矮病毒BYDV RNAI 双T-DNA区载体 无选择标记基因植物 

分 类 号：S435.123[农业科学—农业昆虫与害虫防治]