低能氮离子注入介导沙漠寡营养细菌全基因组突变及定向进化的分子机制

项目名称： 低能氮离子注入介导沙漠寡营养细菌全基因组突变及定向进化的分子机制

项目编号： No.11275164

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 数理科学和化学

项目作者： 毛培宏

作者单位： 新疆大学

项目金额： 80万元

中文摘要： 低能离子注入介导外源DNA的遗传转化已成为远缘、超远缘物种间遗传物质交流的有效手段和方法。本项目通过低能N+注入介导外源多态性DNA片断转化沙漠寡营养细菌，获得其全基因组突变的N+注入参数，建立以沙漠寡营养细菌为模式的离子注入原核微生物全基因组突变的新方法，揭示离子注入导致其全基因组突变的分子机制；利用N+注入介导光能无机自养细菌总DNA和核酮糖二磷酸羧化酶(Rubisco)基因转化处于全基因组突变状态的沙漠寡营养细菌，通过创新的多因子高通量选择性筛选，获得具备固碳固氮功能的重组沙漠寡营养细菌，实现其基因组定向进化的目的；研究重组沙漠寡营养细菌的固碳效率、固氮效率及其协同调控的分子机制。通过本项目的研究，将为二氧化碳的微生物固定和沙漠碳汇工程的研究提供新的思路、方法和理论依据，将为低能离子注入介导的合成生物学的研究与应用提供新的途径。

中文关键词： 氮离子注入；沙漠寡营养细菌；全基因组突变；定向进化；分子机制

英文摘要： The low energy nitrogen ion implantation induced exogenous DNA genetic transformation has become an effective means for distant and super distant interspecies communication of genetic materials. In this project, the exogenous polymorphic DNA fragments were transformed into desert oligotrophic bacteria by low energy nitrogen ion implantation to obtain nitrogen ion implantation parameters of the whole-genome mutation and thus established a new method that used desert oligotrophic bacteria as a model microorganism for whole genome mutation investigation, so as to further reveal the molecular mechanisms of the mutation that caused by ion implantations. Total DNA and Carboxydismutase gene (Rubisco) of photolithotrophical bacteria were transformed into desert oligotrophic bacteria which in whole-genome mutation status by nitrogen ion implantation. Through innovative multi-factor high-throughput selective screening to obtain recombinant desert oligotrophic bacteria with both functions of carbon and nitrogen fixation, and achieved the purpose of the directed evolution of the genome; and further for the investigation of carbon and nitrogen fixation efficiency and the molecular mechanism of the synergistic regulation in recombinant desert oligotrophic bacteria. This Project will provide new ideas, methods and theoretical

英文关键词： nitrogen ion implantation；desert oligotrophic bacteria；whole-genome mutation；directive evolution；molecular mechanism