摘要
脂肪酸氧化酶(LOX)催化不饱和脂肪酸过氧化反应,导致玉米(Zea mays L.)种子储藏品质的下降.为抑制玉米陈化关键酶基因Zmlox-2的表达,依据RNA干扰原理,针对玉米Zmlox-2基因进行克隆,采用重组PCR及酶切连接技术成功构建了Zmlox-2基因的RNA干扰植物表达载体,并将重组载体pRI101-on-Zmlox-2-RNAi通过热激转化法转化到根癌农杆菌中.实验结果表明Zmlox-2基因的RNA干扰表达载体构建正确,这为下一步诱导遗传转化、探索陈化酶基因的干扰表达效果奠定了理论和实验基础.
The peroxidation of unsaturated fatty acid caused by lipoxygenase( LOX) is known to be responsible for the deterioration of storage quality in maize( Zea mays L.) seed. In order to inhibit the expression of key aging enzyme Zmlox-2 gene in maize,Zmlox-2 gene in maize seed was cloned according to the principle of RNA interference( RNAi),and the RNAi plant expression vector containing Zmlox-2 was constructed by means of recombinant PCR and enzyme connection technology. Then,the recombinant vector pRI101-on-Zmlox-2-RNAi was transformed into Agrobacterium tumefaciens via heat shock method. Experimental results show that the RNAi expression vector containing Zmlox-2 gene has been successfully constructed. This research lays a theoretical and experimental foundation for the induction of genetic transformation and the exploration of interference expression effect of aging enzyme gene.
出处
《华南理工大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
2016年第3期136-141,共6页
Journal of South China University of Technology(Natural Science Edition)
基金
吉林省科技发展计划项目(20130102041JC)
吉林省教育厅科学技术研究基金资助项目(吉教科合字[2012]376)~~
