张卫文 陈磊 周杰 马钢 曹旭鹏

摘 要：成功开发了高效、稳定的蓝细菌遗传改造工具。使用该工具可以实现对不同基因进行高表达或低表达；通过理性设计基因组插入位点，实现对蓝细菌内容碳流进行重新分配。该工具正在蓝细菌创建生物合成途径中测试。成功创建了从CO2到异丙醇的生物合成途径，实现利用CO2生物合成异丙醇。突变株生理特性测试及优化正在进行中。构建了针对双组件相应调控蛋白，转录调控因子，以及细胞小RNA等的蓝细菌删除株共273株，以及18株含有不同功能基因的蓝细菌过表达株。使用基于96孔板的底盘检测技术，对构建获得的蓝细菌底盘进行了生物产品耐受性（针对乙醇、丁醇、3羟基丙酸3HP等）以及环境胁迫因子耐受（氮源胁迫、金属离子胁迫、高盐胁迫等）进行了系统的分析，获得多株对于各种胁迫因素以及生物产品发生抗性改变的底盘细胞。确定了一个与集胞藻脂肪酸代谢相关的转录因子，其在脂肪酸代谢网络中的位置及功能的确定还在进一步分析中。完成了两个重要蛋白的晶体结构解析。

关键词：人工合成细胞工厂 光合蓝细菌 底盘细胞

Abstract：A highly efficient and stable genetic transformation tool for cyanobacteria was successfully developped of. High or low expression of different genes can be achieved by use of this tool. A biosynthetic pathway from CO2 to isopropyl alcohol was successfully created. 273 cyanobacteria gene knockout mutants were constructed for the corresponding two- component regulatory proteins， transcription factors， or small RNAs. 18 cyanobacteria overexpression strains containing different functional genes were also constructed. Phenotype analysis was performed and a series of mutants that are tolerant to ethanol， butanol ， 3HP and some environmental stress factors such as nitrogen stress， metal ion stress， high salt stress， were identified. A fatty acid metabolism related transcription factor in Synechocystis was identified. Crystal structure analysis of two important proteins was completed.

Key Words：Synthetic cell factories；Photosynthetic cyanobacteria；Chassis cell

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