人表皮生长因子的原核优势表达与复性  

Optimizing Expression and Renaturation of Recombinant Human Epidermal Growth Factor in E.coli

作　　者：孙卫国[1] 杨栗坤[1] 熊志红[1] 杨秉芬[1] 刘艳华[1] 张灵霞[1] 

机构地区：[1]解放军第309医院结核病研究所全军结核病防治重点实验室,北京100091

出　　处：《生物技术通讯》2015年第5期655-657,共3页Letters in Biotechnology

基　　金：国家传染病防治重大项目(2013ZX-10003-006)

摘　　要：目的:在原核系统中获得具有活性和高表达量的重组人表皮生长因子(rh EGF)。方法:对h EGF编码全序列进行优化,构建原核表达载体p ET-24b-h EGF,在大肠杆菌中表达rh EGF;对rh EGF包涵体进行复性,获得具有较高生物活性的重组蛋白。结果:构建了携带159 bp h EGF基因的表达载体p ET-24b-h EGF,rh EGF在原核系统内得到高表达,重组蛋白相对分子质量为6×103,表达量可占细菌总蛋白的15%,包涵体复性率达90%。结论:对h EGF的编码序列进行了优化,实现了其在原核系统内的高表达,通过复性获得了具有良好生物活性的rh EGF。Objective： To obtain human epidermal growth factor（hEGF） with bioactivity highly expressed in E.co- li. Methods： The gene sequence of hEGF designed by ancillary software Vector NTI suitor 7.0 was synthesized and inserted into plasmid pET-24b to construct pET-24b-hEGF. The pET-24b-hEGF was transformed into E.coli, and hEGF was expressed by induction of IPTG. Then the condition and procedure of renaturation were optimized to improve renaturation ratio and biological activity. Results： The pET-24b-hEGF containing 159 bp modified hEGF gene was constructed. The expression of hEGF about 6 kD in form of inclusion body was up to 25% of to- tal bacterial protein. The renaturation ratio of recombinant hEGF was over 90%. Conclusion： Optimization of gene code of hEGF and renaturation condition have improved expression level of hEGF and its biological activity,

关 键 词：人表皮生长因子 优化表达 包涵体 生物活性 

分 类 号：Q78[生物学—分子生物学]