烟草Ntggpps2基因过表达载体构建及遗传转化

Construction of Ntggpps2 gene overexpression vector and genetic transformation of tobacco

对烟草萜类化合物合成相关的关键基因牻牛儿基牻牛儿基焦磷酸合成酶基因Ntggpps2(Nicotiana tabacum geranylgeranyl diphosphate synthase gene,Ntggpps)进行克隆,并构建了过表达载体p CAMBIA1302-NT2,通过农杆菌介导的遗传转化,成功获得Ntggpps2过表达的烟草转基因植株。qRT-PCR分析发现,在转基因植株的烟草叶片中,Ntggpps2基因的表达水平是野生型的2.2倍,实现了Ntggpps2在烟草中的高表达,为后续研究烟草Ntggpps基因的生物学功能及其萜类物质合成途径的分子机理提供了研究材料与基础。

Nicotiana tabacum geranylgeranyl diphosphate synthase gene（ Ntggpps2） was a key enzyme gene for biosynthesis of the terpenoids and played important roles in plant metabolism and development. In order to study the function of Ntggpps2 in tobacco terpenoid synthesis,the plant overexpression vector of Ntggpps2（ p CAMBIA1302-NT2） was constructed,and Ntggpps2-overexpressing transgenic tobacco plants were obtained by Agrobacterium-mediated genetic transformation. Quantitative RT-PCR was conducted to quantify the transcript accumulation of Ntggpps2 in transgenic plants. The results clearly showed that the expression level of Ntggpps2 was increased by 2. 2 fold in the leaves of transgenic line compared with wild type. The transgenic lines would provide a novel potential material and scientific basis for further biological function analysis of Ntggpps genes and molecular mechanism of terpenoids synthesis in tobacco.