OjCHI真核表达载体的构建及农杆菌的遗传转化

Eukaryotic expression vector construction and Agrobacterium-mediated genetic transformation of OjCHI

查尔酮异构酶(Chalcone isomerase,CHI)是类黄酮生物合成途径中的第二个关键酶,在类黄酮合成与积累过程中发挥着至关重要的作用。笔者以日本蛇根草为材料,在克隆获得其CHI基因(OjCHI)的基础上,将该基因插入由35S启动子控制的真核表达载体pBI121,完成重组表达质粒pBI121-OjCHI的构建,同时利用冻融法将重组质粒成功转入农杆菌GV3101。该实验的完成可为下一步通过转基因技术研究该基因的功能奠定基础。

Chalcone isomerase(CHI)is the second key enzyme in the biosynthesis of flavonoids,which plays an important role during the biosynthesis and accumulation of flavonoids.In this study,we used Ophiorrhiza japonica as materials,and inserted its CHI into eukaryotic expression vector pBI121 driven by 35S promoter to finish the construction of recombinant expression plasmid pBI121-OjCHI based on OjCHI sequence.Then the recombinant expression plasmid was introduced into Agrobacterium tumefaciens strain GV3101 by freeze-thaw method.Overall,the above results will lay an experimental foundation for the further study of OjCHI function by transgenic technology.