两次诱导实现重组大肠杆菌高密度、高表达研究  

Study on the Recombinant Human ApoA-I_Milano High Density and High Expression by Two Temperature-Shifted Induction in Escherichia coli

作　　者：马文峰[1] 庄英萍[1] 郭美锦[1] 丁满生[1] 储炬[1] 张嗣良[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室国家生化工程技术研究中心,上海200237

出　　处：《微生物学通报》2005年第2期54-59,共6页Microbiology China

基　　金：国家高技术研究发展计划("863"计划)(No.2002AA217021);国家重大科技专项资助(No.2002AA2Z3451)

摘　　要：通过不同温度诱导模式对大肠杆菌高密度,高表达的研究,确定两次升温诱导模式实现大肠杆菌高密度、高表达重组人载脂蛋白的目的。实验证实两次诱导成功的避免了乙酸对高密度、高表达的影响,最终发酵的细胞密度OD600达150,蛋白表达量4.8g·L-1,证明两次升温诱导的发酵方法在高密度、高表达外源蛋白上是成功的,从而为基因工程菌规模化生产奠定了基础。The temperature effect on the recombinant protein production formation was investigated in present study. The culture temperature of growth phase is 30℃, and the culture temperature of induction phase was arranged according to three modes. Hign cell-density and high expression culture of E.coli to product recombinant human apolipoprotein A-I Milano by two temperature-shifted induction . Two temperature-shifted induction was carried out high density and high expression recombinant human ApoA-1 Milano. The recombinant protein ApoA-I Milano reached 4.8 g·L -1 with the final cell density of OD 600 150. And the two temperature-shifted induction avoided the acetic acid successfully to the influence of the high density and high expression. Two temperature-shifted induction was viable in high density culture and high expression of heterogenous protein in recombination E.coli.The sduty provides a basic work for production of recombinant ApoA-I Milano in scale.

关 键 词：重组大肠杆菌 ApoA—IMilano 高密度 高表达 两次诱导 

分 类 号：TQ464.7[化学工程—制药化工]