新功能人造生物器件的构建及集成人工细胞的代谢网络改造与系统优化

岳建民

摘 要：该研究采用合成生物学的理念和方法，结合代谢工程的手段，理性设计、构建、优化了埃博霉素在天蓝色链霉菌和紫杉醇、达玛烯二醇在酵母中的异源合成模块。在该研究中，建立了高通量萜类合成途径分析平台。通过基因替换、蛋白表达水平微调控对大肠杆菌MEP途径进行改造，从而构建适合IPP和DMAPP生产的底盘细胞。今年主要的工作为构建大片段DNA合成与组装技术平台，完成了体外全合成全长的简适线粒体DNA分子并完成了测序鉴定。分析了细胞代谢与线粒体形态的变化规律，为日后开展简适线粒体DNA导入后细胞代谢变化分析提供了技术基础。

关键词：合成生物学 代谢工程 紫杉醇 达玛烯二醇 大片段DNA合成

Abstract：Epothilones heterologous synthesis modules in Streptomyces coelicolor， taxadiene and dammarenediol-II heterologous synthesis modules in Saccharomyces cerevisiae were constructed and optimized based on the conception and methods of synthetic biology as well with the methods of metabolic engireening. In our research， a high-throughput screen method for flux-analysis was developed. MEP pathway of E. coli was then engineered by gene replacement and fine-tuning the expression level of key enzymes. In this year， we constructed a de novo large DNA synthesis platform， and accomplished the synthesis of concise mitochondrial DNA and sequencing identification. Furthermore， we investigated the relationship between metabolism switching and mitochondrial morphology， providing the technologies of cellular metabolism detection of cells imported with concise mitochondrial DNA.

Key Words：Synthetic biology；Metabolic engireening；Taxadiene and dammarenediol-II；Large DNA synthesis

阅读全文链接（需实名注册）：http：//www.nstrs.cn/xiangxiBG.aspx？id=49746&flag=1