含绿色荧光蛋白和小鼠T—bet基因双顺反子真核表达载体的构建及其在A549细胞中的表达  

Construction of bi-cistronic eukaryotic vector carrying green fluorescen protein and murine T-bet gene and its expression in AS49 cells

作　　者：檀卫平[1] 刘然义[2] 麦贤弟[1] 吴葆箐[1] 黄绍良[1] 黄文林[2] 

机构地区：[1]中山大学附属第二医院儿科,510120 [2]中山大学肿瘤防治中心实验研究部

出　　处：《现代临床医学生物工程学杂志》2007年第2期109-111,共3页Journal of Modern Clinical Medical Bioengineering

基　　金：广东省科委自然科学基金资助（031668A,06021301A）

摘　　要：目的构建一种带有绿色荧光蛋白（GFP）和小鼠T—bet基因双顺反子的新型真核表达载体pCI—mT—bet—IRES—GFP。方法利用脑炎心肌炎病毒（EMCV）的内部核糖体进入位点（IRES），将GFP的cDNA与小鼠T—bet基因（mT—bet）连接到真核表达载体pCI中，构建含GFP和mT—bet基因双顺反子的重组质粒。结果成功构建了含GFP和mT—bet基因双顺反子的真核表达载体。荧光显微镜下可观察到发绿色荧光的转基因细胞。结论含绿色荧光蛋白和mT—bet基因双顺反子真核表达载体的构建为哮喘基因治疗研究提供了新的生物表达体系。Objective To generate a bi-cistronic eukaryotic vector coexpressing murine T-bet and GFP for facilitating the study of T-bet gene therapy. Methods A 600 bp IRES from encephalomyocarditis virus and 700 bp GFP were trimolecularly ligated into EcoR I site of eukaryotic vector pCI. An approximately 1.7 kb murine T-bet cDNA segment was then ligated into Xho I site of pCI. Result The bi-cistronic plasmids coexpressing murine T-bet and GFP was identified by restriction enzymes map. Conclusion The murine T-bet cDNA eukaryotic expression vector pCI-mT-bet-GFP is successfully constructed, which can provide a new expressive system for the research of gene therapy in asthma.

关 键 词：小鼠T—bet基因 绿色荧光蛋白 真核表达载体 

分 类 号：R[医药卫生]