作　　者： ; ; ; ; ; ;

机构地区： 厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室

出　　处： 《厦门大学学报（自然科学版）》 2003年第4期410-415,共6页

摘　　要： 以本室构建的原核表达载体pTO-T7为基础载体,PCR合成ompT引导序列,插入该载体多克隆位点上游,构建了分泌型原核表达载体pTO-OT。将2个外源基因克隆至pTO-OT,2个重组质粒在大肠杆菌中均得以高效表达,表达产量在25％～34％之间。Western blot分析证实了融合蛋白可被大肠杆菌信号肽酶有效地切割,并具有良好的免疫学活性。对重组表达菌株的连续传代实验证实了该表达载体具有良好的表达稳定性,显示了其在基因工程中的应用价值。 A prokaryotic secretion expression vector, pTO-OT, was constructed based on expression vector pTO-T7 by inserting ompT leader sequence into the MCS of pTO-T7. Two foreign genes were cloned into pTO-OT and could be expressed efficiently in E. coli. Ratios of each recombinant protein to total bacteria proteins varied from 25% to 34%. Western blot analysis on the two recombinant proteins suggested that they could be cut by signal peptidase in E. coli, and the mature proteins were immunoactive. The consecutive culture of recombinant engineering strains showed pTO-OT has excellent expression stability, which suggested pTO-OT was a practical vector for genetic engineering.

关 键 词： 原核表达载体 基因工程 基因克隆 基因表达 载体构建 分泌型

领　　域： [生物学]