新功能人造生物器件的構建及集成人工細胞的代謝網絡改造與系統優化

岳建民

摘 要：該研究采用合成生物學的理念和方法，結合代謝工程的手段，理性設計、構建、優化了埃博霉素在天藍色鏈霉菌和紫杉醇、達瑪烯二醇在酵母中的異源合成模塊。在該研究中，建立了高通量萜類合成途徑分析平臺。通過基因替換、蛋白表達水平微調控對大腸桿菌MEP途徑進行改造，從而構建適合IPP和DMAPP生產的底盤細胞。今年主要的工作為構建大片段DNA合成與組裝技術平臺，完成了體外全合成全長的簡適線粒體DNA分子并完成了測序鑒定。分析了細胞代謝與線粒體形態的變化規律，為日后開展簡適線粒體DNA導入后細胞代謝變化分析提供了技術基礎。

關鍵詞：合成生物學 代謝工程 紫杉醇 達瑪烯二醇 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

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