CAJ | 학술논문

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 dwarfvirus,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.
RNAi계쌍련RNA유도동원mRNA강해,도치특정기인표체침묵적일충현상.RNAi기술시통과기인공정방치식물병독병적최가도경.유대맥황왜병독(Barely yellow dwarfvirus,간칭BYDV)인기적화곡류황왜병발병면적화위해정도재아국정가중추세.BYDV-GAV시당전류행적대맥황왜병독주계,본문침대BYDV-GAV복제매기인서렬,구건능재세포내전록형성발잡RNA쌍련결구적표체합,병장기치우중간재체pVec8-2b적T-DNA구,pVec8-2b적령일T-DNA구함유hpt선택보고기인표체합.구쌍T-DNA구적병독복제매기인발잡RNA고효표체재체이도입근암농간균,가용우유도식물RNAi,창제무선택표기기인적항대맥황왜병전기인작물.
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 hygromyein 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.Fig6,Ref21