我们已率先克隆人参皂苷转运蛋白候选PgPDR3基因，且在人参和爪蟾卵母细胞表达证实其转运人参总皂苷功能。但PDR3表达蛋白对皂苷组分的特异性转运和皂苷生物合成的调控作用及机制尚不清楚。本研究拟建立PDR3超表达的酵母系并使用皂苷组分喂饲，LC-MS定量测定上述酵母细胞中皂苷含量变化，同时应用SPR和ITC法体外鉴定酵母PDR3蛋白与皂苷组分的互作，进而阐释PDR3蛋白特异性转运皂苷的功能；建立PDR3超表达及CRISPR/Cas9系统敲除的人参细胞和发根系，应用功能组学技术比较分析其转录组表达与皂苷生物合成通路代谢组改变的关系，以阐明PDR3对皂苷生物合成的调控作用与机制；再建立PDR3与皂苷生物合成主途径DDS超表达、分支代谢βAS/CAS敲除的人参发根系，从转录组及代谢组水平探究PDR3协同调控在皂苷生物合成与积累中的作用。最终为实现达玛烷型人参皂苷代谢工程生产与利用提供新的理论依据。

The PgPDR3 gene involved in ginsenosides transport has first been cloned and its functional expression analysis in Xenopus oocyte and Panax ginseng cells showed PDR3 transporter gene played function in the transport of total ginsenosides. However PDR3 function involved in ginsenoside components-specific transport and ginsenosides biosynthesis is unclear. Based on these results, Yeast cells (ABC transporter genes knockout) harboring PDR3 gene will be used as ginsenosides precursor and its metabolic components feeding test to quantitatively analyze the changes of the ginsenosides using LC-MS/MS. The specific recognition and interaction between ginsenosides and PDR3 transporter will be further identified by means of the surface plasmon resonance (SPR) and the isothermal titration calorimetry (ITC) to clarify the function of PDR3 specific recognition, transport of ginsenoside components. Using transcriptome sequencing and LC-MS metabolome techniques to comparatively analyze the relationship between the expression of transcriptome and changes of the ginsenoside biosynthesis metabolomic pathway in ginseng cell and hairy root with PDR3 overexpression and CRISPR/Cas9 system knock-out, and then the function of PDR3 in regulating ginsenoside biosynthesis will be further elucidated. To further explore the function played in ginsenoside biosynthesis and accumulation by PDR3 and ginsenoside biosynthases coordinated expression, PDR3 overexpression will be combined with the overexpression of DDS that is considered as the key enzyme of dammarane-type ginsenoside biosynthesis and also with CRISPR/Cas9 system knock-out of the key enzyme genes including β-AS and CAS in the branch metabolism pathway of ginsenoside biosynthesis. Finally, this study will provide a theoretical base and a new strategy in improving the dammarane-type ginsenoside yield through the coordinated regulation of transport and biosynthesis metabolism.