融合蛋白表达载体的构建及在大肠杆菌中的表达  

Reconstruction and Expression of Regucalcin Fusion Protein in E. coli

作　　者：李莉[1] 高秀来[1] 宋一志[1] 陆涛[1] 景鹏[1] 

机构地区：[1]首都医科大学基础医学院解剖学教研室

出　　处：《首都医科大学学报》2006年第6期788-791,共4页Journal of Capital Medical University

摘　　要：目的构建人类regucalc in重组原核表达载体。方法用RT-PCR的方法从人类肝癌细胞系HepG2中扩增出regucalc in基因cDNA序列,应用T克隆载体及pET-32 a(+)表达载体后,将质粒转化进入原核表达菌株进行表达。结果表达产物经SDS-PAGE电泳分析后,在45 000处有一明显表达条带,与理论结果相符。结论成功构建融合表达载体Pet-regucalc in,并在原核细胞中表达出人类regucalc in-H is融合蛋白。Objective To construct the prokaryotic expression vector of the human regucalcin gene. Methods To get the human regucalcin cDNA from human live cancer HepG： cells with RT-PCR, and then inserted the fragment of regucalcin into pMD-18 simple T vector. The amplified products were cloned into the prokaryotic expression vector pET-32a（ ＋ ）. Results There was one obvious band at the position of relative molecular weight 45 000 after SDS-PAGE analysis, and it was equivalent to the expected value. Conclusion To construct the recombinant fusion expression vector and to express the fusion protein in E. coli successfully.

关 键 词：RT—PCR REGUCALCIN 基因表达 融合蛋白 

分 类 号：Q782[生物学—分子生物学]