摘要
为了确定出rhTNFα最佳表达条件 ,实验利用正交试验设计方法 ,对rhTNFα工程菌的进行发酵培养和 4 2℃热诱导 ,此后借助蛋白凝胶电泳分析软件对各组样品的凝胶电泳图片定量分析 ,利用方差分析对影响TNF基因工程菌对目的产物表达的几个重要的条件进行了优化 ,并最终确定出 pH7.5、培养温度 2 7℃、培养时间 4 .5h、诱导时间 5h的最佳表达条件。在该条件下进行中试发酵TNF的表达产物占到了菌体总蛋白的 6 0 %左右 ,且大部分以可溶形式存在。
To seek out optimal expression conditions of recombinant human TNF α. Designing this experiment with orthogonal test,we cultured engineering bacteria strain and induced the expression of rhTNF α at 42℃, then run SDS PAGE. The expression level of rhTNF α was quantified by analysis software,and several important expression conditions were optimized by variance analysis of the data. The optimal expression condition of recombinant human TNF α is pH 7.5 ,culturing temperature 27℃, culturing time 4.5h, inducing time 5h,and the products were obtained with about 60% yield of total bacterial proteins in pilot scale fermentaion.Low culture temperature is favorable to expression of rhTNF α and can reduce formation of inclusion bodies on condition that is does not limit growth of E.coli.
出处
《微生物学免疫学进展》
2003年第2期12-15,共4页
Progress In Microbiology and Immunology
关键词
TNF
表达条件
正交试验设计
中试发酵
Recombined human tumor necrosis factor α (rhTNF α)
Expression condition
Orthogonal experimental design
Pilot scale fermentation
