设计构建人工细胞生产萜烯类天然药物具有重大研究价值，然而其MVA路径通量低是限制因素之一。研究表明非必需基因的全局调控可有效地提高MVA路径的通量。申请者设想通过诱导CRE-LoxP进行SCRAMBLE基因重排，从全局扰动来提高MVA路径通量。主要研究：1）以化学合成V号染色体人工合成酿酒酵母为底盘细胞，对非必需基因进行重排，快速构建多靶点、多种扰动方式高效调控MVA路径通量的人工细胞；2）构建β-胡萝卜素模块作为MVA通量的指示，通过差异表型解析潜在基因靶点及其对MVA路径关键基因、蛋白和代谢物，精准指导高MVA路径通量底盘的重构；3）基于获得的新知识，重构人工细胞，进行表征及扰动信息修正。该研究为人工细胞生产天然产物进行全局精准调控提供了新的思路和解决办法。

Design and construction of artificial cell factories for natural drug-terpenes production is of great importance. However, inadequate MVA pathway flux is one of the limiting factors. It is reported that global manipulation of non-essential genes can improve the flux of MVA pathway effectively. The applicant proposes to improve MVA pathway flux through SCRAMBLE gene re-arrangement induced by CRE-LoxP. This project intends to mainly focus: 1) using artificial Saccharomyces cerevisiae with V chromosome from chemical synthesis as chassis, an artificial cell with high MVA pathway flux will be built rapidly through multi-targets and multiple disturbance pattern by non-essential genes rearrangement; 2) Using beta-carotene module as reporter for MVA pathway flux, potential gene targets in different phenotype and their disturbance information on key genes, proteins and metabolites in MVA pathway will be uncovered, which will be applied for precisely guiding the reconstruction of chassis with high MVA pathway flux; 3) Based on the knowledge we got, through reconstruction of artificial cells, characterization and disturbance information correction will also be done. This study provides a new insight and solution for global and precise manipulation on artificial cells for natural products production.