目的:构建带有特异人端粒酶逆转录酶(hTERT)启动子及猪α半乳糖基转移酶(α1,3GT基因)的真核表达载体,使目的基因选择性地在肿瘤细胞中表达α半乳糖(α-Gal)抗原表位,不影响其它正常组织细胞,通过人体内预存的天然抗α-Gal抗体激活超急性排斥反应从而准确的杀伤肿瘤细胞。 方法:通过基因克隆的方法构建带有绿色荧光蛋白的异源α1,3GT基因的pEGFP-α1,3GT重组质粒及pEGFP-hTERT-α1,3GT重组质粒。测序鉴定构建成功的重组质粒通过脂质体法转染A549(人肺腺癌细胞系)及电穿孔法转染原代HUVEC(人脐静脉内皮细胞),通过荧光显微镜、观察目标蛋白的定位,流式细胞仪观察重...Objective :To construct the p EGFP--h TERT-α 1,3GT plasmid which can selectivity express α-Gal antigen in the tumor cells nor normal tissue and cells. To induce hyperacute rejection by antiα-Gal natural antibody for precisely kill the tumor cells. Methods: To construct and identify the p EGFP-α 1,3GT and p EGFP--h TERT-α 1,3GT plasmids by the methods of gene cloning. To transfect A549cell line by...学位:医学硕士院系专业:医学院临床医学系_内科学学号:2452007115255