用原核翻译增强子提高人成骨生长肽基因在大肠杆菌中的表达  

Improving expression of human osteogentic growth peptide gene in E.coli.with the enhance of protein synthesis initiation

作　　者：周凌云[1] 刘兴汉[1] 余琼[1] 

机构地区：[1]哈尔滨医科大学生物化学与分子生物学教研室,黑龙江哈尔滨150086

出　　处：《哈尔滨医科大学学报》2003年第5期379-382,共4页Journal of Harbin Medical University

基　　金：国家自然科学基金资助项目 ( 3 9980 0 0 4)

摘　　要：目的　用原核翻译增强子提高人成骨生长肽基因在大肠杆菌中的表达。方法　将原核翻译增强子序列插入到质粒pRIT 2T启动子的下游 ,再与成骨生长肽融合蛋白基因重组。热诱导表达融合蛋白 ,经亲和层析、FactorXa酶切、凝胶过滤层析得到纯化的成骨生长肽。结果　提高了成骨生长肽基因在大肠杆菌中的表达水平 ,使成骨生长肽融合蛋白的表达量超过了大肠杆菌蛋白总量的 5 0 %。结论　原核翻译增强子可提高外源基因在大肠杆菌中的表达。插入了原核翻译增强子的pRIT 2T质粒可作为高效表达融合蛋白的通用质粒载体。Objective To improve expression of human osteogentic growth pepti de (OGP) gene in E.coli.with the prokaryotic translational enhancer. Methods The OGP gene was cloned into the plasmid pRIT-2T,in front of which promoter had been inserted the sequence of prokaryot ic translational enhancer. The recombinant plasmid was induced by heat. T he fusion protein was purified through affinity chromatography and cleaved by fa ctor Xa,and OGP was isolated by gel filtration. Results Human OGP gene in E.coli. was expressed effectively. Conclusion The plasmid pRIT-2T inserted the enhance can be used as the current vecter expressing fusion protein ef fectively.

关 键 词：原核翻译增强子 人成骨生长肽 基因表达 大肠杆菌 基因重组 

分 类 号：Q516[生物学—生物化学]