谷氨酸棒状杆菌新型诱导启动子的研究

Study of the New Inducible Promoter in Corynebacterium Glutamicum

以丰富和完善谷氨酸棒状杆菌的表达元件、筛选适合工业化应用的新型诱导启动子为研究目标,借助绿色荧光蛋白GFP为报告基因,构建启动子筛选工具质粒,在谷氨酸棒杆菌中表达,筛选出三种高效表达的诱导启动子.通过检测绿色荧光蛋白表达强度可知:丙酸启动子诱导表达强度最大,葡萄糖酸启动子次之,蔗糖启动子较弱;丙酸启动子最适诱导浓度为60 ug/m L、葡萄糖酸启动子2 mg/m L、蔗糖启动子2 mg/m L,表达强度比无诱导对照组分别增加4.39、3.86、2.74倍.

To enrich the expression elements of Corynebacterium glutamicumand screen out new available inducible promotets in industrial application, the article usesGreen fluorescent protein （GFP）as report system to construct promoter selecting articles and expresses them in Corynebacterium glutamicum. Three highly expressed inducible promoters are selected. By measuring expression levelof Green fluorescent protein, we demonstrate that propionate promoter is the highest of all in induction intensity, gluconate promoter comes the second, and sucrose promoter is the lowest. The propionate concentration of only 60 ug/mL is found to be sufficient for full induction（increasing 4.39 fold）of its native target.Adding 2mg/mL gluconate and sucrose respectively, the expression intensity reaches its maximal induction, increasing 3.86 fold and 2.74 fold than its native target respectively.