论文部分内容阅读
目的:克隆人Egr-1基因的启动子,插入荧光素酶报告基因载体中,并检测电离辐射对其活性的影响。方法:采用PCR技术从人乳腺癌细胞系MCF-7基因组中扩增出Egr-1启动子,将其克隆到pGL3-basic载体中;将重组质粒转染人肿瘤细胞,测定Egr-1启动子在不同辐射条件下转录活性的改变。结果:成功构建了Egr-1启动子的荧光素酶报告基因;在不同剂量的γ射线照射后,Egr-1的启动子活性均明显高于未照射组;在同一剂量照射后48 h,Egr-1的启动子活性达峰值。结论:本实验构建的Egr-1启动子具有辐射激活的功能,为进一步研究放射-基因治疗奠定了基础。
OBJECTIVE: To clone the promoter of human Egr-1 gene into luciferase reporter gene vector and detect the effect of ionizing radiation on its activity. Methods: The Egr-1 promoter was amplified from human breast cancer cell line MCF-7 by PCR and cloned into pGL3-basic vector. The recombinant plasmid was transfected into human tumor cells and the expression of Egr-1 promoter Changes in transcriptional activity under different irradiation conditions. Results: The luciferase reporter gene of Egr-1 promoter was constructed successfully. The promoter activity of Egr-1 was significantly higher than that of non-irradiated Egr-1 cells after irradiated with different doses of γ-rays. At 48 h after irradiation with the same dose of Egr -1 promoter activity peaked. Conclusion: The Egr-1 promoter constructed by this experiment has the function of radiation activation, which lays the foundation for further study of radiation-gene therapy.