融合蛋白表达载体的构建及在大肠杆菌中的表达

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。

Reconstruction and Expression of Regucalcin Fusion Protein in E.coli

Objective To construct the prokaryotic expression vector of the human regucalcin gene.Methods To get the human regucalcin cDNA from human live cancer HepG_2 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.