PttGA20-氧化酶基因dsRNA抑止载体的构建

[目的]通过基因工程手段抑止赤霉素调控基因PttGA20-氧化酶基因的表达,从而抑制植物高生长和节间伸长,达到培育矮化植株的目的。[方法]依据RNAi原理,设计引物扩增正义及反义PttGA20ox片段插入pBI121载体CaMV35S启动子下游区域,并在正义和反义片段间隔区插入茎环结构GUS基因片段,将npⅡt标记基因替换为抗除草剂基因bar,构建dsRNA抑止载体。[结果]所构建的抑止载体经经不同内切酶酶切鉴定后均可释放出与目的条带大小相同片段,表明已成功构建PttGA20-氧化酶基因dsRNA抑止载体。[结论]该研究为培育矮化植株提供了新的途径。

RNAi Plasmid Construction of PttGA 20-oxidase Gene

[Objective] Genetic Engineering technology was used to regulate the expression of PttGA20-oxidase gene thus restrained plant height growth and internode elongation for cultivating dwarfed plant.[Method] Based on the RNAi principle,the gene specific sequences of PttGA20-oxidase in the antisense and sense orientations interrupted by a gene sequence from GUS were cloned into a binary vector pBI121.The selection marker gene npt Ⅱ was replaced with bar gene to RNAi plasmid.[Result] After undergone different endonuclease restrictions,the constructed constraint vector released different segments whose sizes were similar to that of target segment,which demonstrated that the RNAi plasmid of PttGA20-oxidase gene was successfully constructed.[Conclusion] The experiment provided a new way for culturing dwarfed plant.