转TNF-α基因大肠杆菌培养条件的优化  

Optimization for Cultivation Conditions of Transgenic Escherichia coli Harbering TNF-α Gene

作　　者：李丹[1] 韩睿[1] 李轩[1] 施定基[1] 张芃芃[2] 宋东辉[1] 王学魁[1] 

机构地区：[1]天津科技大学海洋科学与工程学院,天津300457 [2]中国科学院植物研究所,北京100093

出　　处：《生物技术》2008年第2期60-64,共5页Biotechnology

基　　金：天津市科委重点攻关项目("蓝藻基因工程制备TNF-α口服剂的研究和开发";043182711)

摘　　要：目的:优化转pMD-489-TNF大肠杆菌的发酵条件,提高菌体产量及TNF-α产率。方法:通过单因素及正交实验,对影响转基因大肠杆菌生长及TNF-α表达的培养液成分、诱导剂浓度、培养温度等工艺条件进行了优化。结果:M9+LB培养基成分的最优配比为3%葡萄糖、2%蛋白胨、4%酵母粉和1%NH4Cl,诱导剂IPTG浓度为0.5mmol/L,诱导时间4h,培养液初始pH是7.0,于37℃培养。结论:优化后的培养条件促进了菌体的生长和TNF-α的表达,菌体干重和TNF-α表达率比没有经过优化时升高了1.34倍和4.11倍,TNF-α产率从0.113%提高到0.479%,提高了324%。Objective：In order to stimulate the growth and the expression of Tumor Necrosis Factor-α（TNF-α） in Escherichia coli,fermentation conditions of transgenic E.coli were optimized.Method：Single factor experiment and orthogonal experiment were conducted to study fermentation conditions such as composition of culture,concentrations of IPTG and fermentation temperature etc.which affect the growth of E.coli and expression of TNF-α.Result：By range analysis and variance analysis,the optimum fermentation conditions were obtained as followed：the glucose 3%,the tryptone 2%,the yeast 4%,the NH4Cl 1%,the concentration of TPTG 0.5mmol/L,revulsive time 4h,initial pH 7.0,fermentation temperature 37℃.Conclusion：By optimizating fermentation conditions,the yield of E.coli and the expression efficiency of TNF-α could be raised 1.34 fold and 4.11 fold respectively,the yield of TNF-α increased from 0.113% to 0.479%,that had been raised 324% as compared with the original yeild.

关 键 词：肿瘤坏死因子Α 大肠杆菌 优化培养 基因工程 

分 类 号：Q813.11[生物学—生物工程]