应用珠蛋白启动子驱动荧光蛋白在K562细胞中表达  

Expression of fluorescence protein in K562 cells driven by erythroid-specific promoter

作　　者：谢书阳[1] 王萍玉[2] 姚宝洪[1] 焦飞[1] 王桂兰[1] 杨成[3] 

机构地区：[1]滨州医学院生物化学与分子生物学教研室,滨州市256603 [2]滨州医学院卫生学教研室 [3]滨州医学院中心实验室

出　　处：《滨州医学院学报》2008年第1期8-10,共3页Journal of Binzhou Medical University

基　　金：山东省教育厅基金(106L01);滨州医学院大学生科技创新活动基金项目

摘　　要：目的应用红系特异性启动子驱动荧光蛋白(绿色荧光蛋白GFP和红色荧光蛋白RFP)在血液病细胞K562中表达,以验证克隆所得红系特异性启动子的有效性。方法应用PCR的方法扩增红细胞系特异性启动子,构建该启动子驱动GFP基因表达载体和驱动RFP基因表达载体。然后转染K562细胞,用荧光显微镜、RT-PCR分析基因表达的情况。结果荧光显微镜示在K562细胞中可见较强的绿色荧光和红色荧光表达,RT-PCR进一步验证了GFP和RFP能在K562细胞中有效的表达。结论克隆的红细胞系启动子是有效的,能有效地启动目的基因在血液病细胞中表达,为下一步进行血液病基因治疗提供科学的资料。Objective Studing the effect of erythroid-specific promoter can effectively drive expression of green/red fluorescence protein（GFP/RFP） in K562 cells. Methods Erythroid-specific promoter（ β globin promoter） was amplified by PCR from human genome DNA, which was cloned into pcDNA vector to drive GFP/RFP gene expression. Then the vectors were transfected into K562 cells to prove the effect of the promoter by using fluorescence microscopy and RT-PCR. Results Fluorescence microscopy showed that GFP/ RFP can strongly express in K562 cells. And the mRNA was also shown to express effectively by RT-PCR analysis. Conclusion The erythroid-specific promoter can obviously drive gene expression in K562 cells, which is valuable for gene therapy to leukemia as well as other hematology diseases.

关 键 词：红细胞系特异性启动子 荧光蛋白 基因表达 

分 类 号：Q75[生物学—分子生物学]