巨噬细胞特异性启动子调控的真核表达载体的构建及靶向性研究  

Construction and targeting study of eukaryotic expression vector modulated by a macrophage-specific promoter

作　　者：文阳安[1] 郭金英[1] 余晓林[1] 岑东[1] 刘靳波[1] 陈彦[2] 张健[3] 涂植光[1] 

机构地区：[1]重庆医科大学医学检验系,临床检验诊断学省部共建教育部重点实验室,重庆市重点实验室,重庆400016 [2]西南大学医院,重庆400715 [3]重庆医科大学附属第一医院骨科,重庆400016

出　　处：《细胞与分子免疫学杂志》2008年第5期441-443,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30600790)

摘　　要：目的:构建巨噬细胞特异性启动子调控的靶向巨噬细胞的真核表达载体。方法:PCR方法合成巨噬细胞特异性启动子,并将其插入带有绿色荧光蛋白(GFP)基因的真核表达载体pEGFP-N1中,替代CMV启动子。利用脂质体转染法,将其与红色荧光蛋白真核表达载体(pERFP-N1)共转染巨噬细胞和非巨噬细胞,通过荧光显微镜观察GFP和RFP在不同细胞中的表达水平来鉴定启动子的靶向性。结果:成功构建了巨噬细胞特异性启动子调控的真核表达载体,并且能在巨噬细胞中高效特异性地表达报告基因。结论:构建的靶向巨噬细胞的真核表达载体能提高目的基因的表达特异性,为提高基因治疗胞内菌感染的靶向性及降低毒副作用奠定了实验基础。AIM： To construct a macrophage-specific eukaryotic expression vector and to investigate its expressing specificity. METHODS： Macrophage-specific promoter was synthesized using PCR, and substituted the CMV promoter of eukaryotic expression vector pEGFP-N1 to construct a recombinant vector （ pSP-GFP）, which were cotransfected with pERFP-N1 vector into different cell lines. The green fluorescent protein （GFP） and red fluorescent protein （RFP） was observed by fluorescence microscopy, and the target specificity of macrophage-specific promoter was judged by comparison of expressed level of GFP and RFP in various cell lines. RESULTS： The macrophage-specific eukaryotic expression vector was successfully constructed, which showed strong activity and specificity only in macrophage cells. CONCLUSION： The constructed pSPGFP vector was macrophage-specific, which are potential to targeted gene therapy of intracellular bacterial infection.

关 键 词：巨噬细胞 细胞特异性 启动子 真核表达载体 胞内菌感染 基因治疗 

分 类 号：R392.12[医药卫生—免疫学]