含不同启动子的GFP载体在不同种属的胚胎干细胞系中的表达效率  

Effect of Different GFP Expression Vectors with Different Promotors on Expression Efficiency in Mouse Embryonic Stem Cells and Human Embryonic Stem Cells

作　　者：陈红[1] 钱坤[2] 张苏明[3] 

机构地区：[1]华中科技大学同济医学院附属同济医院康复医学科,湖北武汉430030 [2]华中科技大学同济医学院附属同济医院生殖医学中心,湖北武汉430030 [3]华中科技大学同济医学院附属同济医院神经科,湖北武汉430030

出　　处：《武汉大学学报（医学版）》2007年第2期134-137,146,I0003,共6页Medical Journal of Wuhan University

基　　金：国家自然科学基金项目(编号:30370505;30600188)

摘　　要：目的:对比含不同启动子的绿色荧光蛋白(GFP)表达载体在hESC及小鼠胚胎干细胞(mESC)的表达效率,为探索ESC及其衍生细胞移植在体内的存活、迁移、分化及整合提供有力的细胞模型。方法:阳离子脂质体转染hESC及mESC ES-D3,荧光显微镜下观察阳性克隆,流式细胞术(FCM)计算不同载体pCX-hrGFP、pIRES-hrG-FP在不同种属细胞中的表达效率。结果:两种载体在mESC的表达效率分别为pCX-hrGFP(90±2.5)%,pIRES-hrGFP(0.67±0.02)%,两组间比较差异有统计学意义(P<0.05)。在hESC的表达效率分别为pCX-hrGFP(0.8±0.1)%,pIRES-hrGFP(0.62±0.08)%,两组间比较有统计学差异(P<0.05)。pCX-hrGFP在mESC及hESC间的表达效率差异有统计学意义(P<0.01),pIRES-hrGFP在mESC及hESC间的表达效率差异无统计学意义(P>0.05)。结论:①CBA启动子引导的GFP表达效率高于CMV启动子。②同一载体在不同种属细胞内表达效率不同。Objective： To compare effect of different GFP expression vectors with different promoter on expression efficiency in hESC and mESC and offer a tool for exploring the survival, transferring, differentiation and integration of ESC and its derivatives in vivo. Methods： The hESC and mESC lines were transfected with lipofectamine 2000, positive colonies were detected under fluorescent microscope, and expression efficiency were analized and compared between different transfect vectors such as pCX-hrGFP, pIRES-hrGFP in different cell lines. Results： Expression efficiency of pCX-hrGFP and pIRES-hrGFP was （90 ±2.5）% and （0.67±0.02） % separately in mESC and they had statistically difference （P〈0.05）. It was （0. 8±0. 1）% and （0.62±0. 08）% in hESC seperately, which had statistically difference （P〈0.05）. There was significantly difference in expression effiency of pCX-hrGFP between mESC and hESC （P〈0. 01） ,while it had no difference of plRES-hrGFP between the two groups （P〉0. 05）. Conclusion： ① Expression efficiency of GFP driven by CBA promoter is highter than by CMV promoter. ② Expression efficiency of the same expression vector is different in cells from different species.

关 键 词：胚胎干细胞 基因转染 表达效率 

分 类 号：Q813[生物学—生物工程]