谷氨酸棒杆菌是一种广泛用于各种细胞代谢产物生产的重要工业微生物，同时，也是一种理想的重组蛋白生产宿主菌。目前使用的原核重组蛋白生产系统都有各自的缺陷。本项目通过构建使用T7-lacO调控区的分泌型表达载体pAU-4，以及染色体上整合大肠杆菌T7 RNA聚合酶基因和LacI阻遏蛋白基因的工程宿主菌株C. glutamicum XU1，优化重组蛋白生产条件，建立一套谷氨酸棒杆菌高效严谨基因表达系统，为重组蛋白高效生产提供一条非常有应用前景的新途径。基于RecA同源重组的基因打靶技术已不能完全满足谷氨酸棒杆菌功能基因组学、代谢网络及其调控和代谢工程研究的需要。本项目通过在谷氨酸棒杆菌中构建Red同源重组系统pAU-6以及FLP位点特异性重组系统pAU-8，优化打靶条件，建立一套简单高效的新型谷氨酸棒杆菌基因打靶系统，为谷氨酸棒杆菌的基础和应用研究提供一套简单高效的基因打靶新方法。

Corynebacterium glutamicum is not only an important industrial microorganisms that has been widely used in the production of a variety of cellular metabolites, but a desired host used to produce recombinant protein. The existing recombinant protein production systems have their own shortcomings. In this project, by constructing secretory expression vector pAU-4 harboring T7-lacO regulation region and engineering host strain C. glutamicum XU1 that Escherichia coli T7 RNA polymerase gene and LacI repressor protein gene were integrated on chromosome, optimizing recombinant protein production conditions, a high-efficient and stringent Corynebacterium glutamicum gene expression system would be estabished, which would provide a very promising way for the high-efficient production of recombinant proteins. Gene targeting technology based on RecA homologous recombination has not fully meet the needs of functional genomics，metabolic networks and their regulation and metabolic engineering studies for C. glutamicum. In this project, by constructing plamid pAU-6 harboring the Red homologous and FLP site-specific rocombination systems for C. glutamicum and optimizing gene targeting conditions, a simple and high-efficient new gene targeting system would be estabished, which would provide a new method to the research for C. glutamicum.