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,
出处
《生物技术通讯》
CAS
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)
关键词
人表皮生长因子
优化表达
包涵体
生物活性
human epidermal growth factor
optimizing expression
inclusion body
biological activity