ProZZ-EGFP融合蛋白基因在大肠杆菌中分泌表达条件的优化  

Optimization of ProZZ-EGFP fusion protein secreted expression in E.coli

作　　者：唐金宝[1] 李万忠[1] 吉爱国[2] 孙利民[2] 朱鹏[2] 

机构地区：[1]潍坊医学院基础部药学教研室,山东潍坊261042 [2]山东大学药学院,山东济南250012

出　　处：《中国生化药物杂志》2007年第1期37-39,共3页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的优化大肠杆菌分泌表达ProZZ-EGFP融合蛋白基因的培养条件。方法采用摇瓶培养,在液体培养基中加入终浓度不同的蔗糖、Triton X-100和甘氨酸,诱导大肠杆菌周质腔内蛋白质“泄漏”到液体培养基中,利用ProZZ-EGFP浓度-荧光强度标准曲线快速检测培养基中目的蛋白质浓度。结果利用大肠杆菌HB101表达ProZZ-EGFP融合蛋白,在培养基中含有终浓度1%Triton X-100及1%甘氨酸,可使ProZZ-EGFP在培养液的分泌表达量提高6倍。结论仅在培养基中加入几种物质即可提高ProZZ-EGFP融合蛋白的分泌表达量,简单易行。Purpose To enhance the yield of the ProZZ-EGFP excretion into culture media. Methods The bacteria were grown in shaking flasks, when A600nm at 0.6, the cell culture was added with sucrose, glycine or Triton X-100 at different concentrations that affect the excretion of the fusion protein into culture media. By detecting the fluorescence intensity of the supernatant, we can assess the level of excretion of ProZZ-EGFP into the liquid cultures easily and quickly. Results ProZZ-EGFP secreted into culture media with 1% glycine and 1% Triton X-100 is 6-fold higher than that without the two chemicals. Conclusion This study provides a practical, large-scale method for more efficient production of the ProZZ-EGFP fusion protein in E. coli by using glycine and Triton X-100.

关 键 词：ProZZ-EGFP 大肠杆菌 分泌表达 优化 

分 类 号：R392-33[医药卫生—免疫学]