ME: Collaborative Research: Metabolic Engineering of Hairy Roots for Alkaloid Production

本项目的目的是提高吲哚生物碱在一个良好的模式植物组织培养系统，C。罗斯 总体目标是确定遗传操作的一些关键酶的吲哚和非甲羟戊酸萜类化合物途径通量通过吲哚生物碱合成途径的代谢效应，特别强调初级和次级代谢之间的相互作用，最终生物碱生产力。 本研究的具体目标是：1）构建和鉴定在最近鉴定的诱导型启动子系统控制下过表达关键酶如ORCA 3、G10 H、DXS、DXR和AS-β的转基因毛状根系; 2）开发基于NMR的代谢通量图谱的方法; 3）对这些第一代转基因系进行代谢表征; 4）开发允许引入多个基因的共转化技术; 5）构建和表征表达基因的策略组合的第二代转基因系，如从上述第一代转基因系的结果确定的。

The objective of this project is to improve the productivity of indole alkaloids in a well-characterized model plant tissue culture system, C. roseus. The overall goals are to determine the metabolic effects of genetic manipulations of some key enzymes in the indole and non-mevalonate terpenoid pathways on flux through the indole alkaloid synthesis pathways, with special emphasis on the interplay between primary and secondary metabolism on the final alkaloid productivity. The specific objectives are to: 1) construct and characterize transgenic hairy root lines overexpressing key enzymes, such as ORCA3, G10H, DXS, DXR and AS-beta under the control of a recently characterized inducible promoter system; 2) develop methods for NMR- based metabolic flux maps; 3) perform metabolic characterization of these first generationtransgenic lines; 4) develop techniques for co-transformation that allow the introduction ofmultiple genes; 5) construct and characterize a second generation of transgenic lines expressing strategic combinations of genes as determined from the results of the first generation transgenic lines above.