摘要
目的:构建DEK的pcDNA3-Flag表达载体,研究其对抑癌基因p53启动子活性的影响。方法:以乳腺文库为模板,PCR扩增DEK编码序列,克隆到pcDNA3-Flag载体,构建成pcDNA3-Flag-DEK,转染293T细胞,Western印迹鉴定pcDNA3-Flag载体介导的DEK的表达,萤光素酶报告基因活性实验研究DEK对p53启动子活性的影响。结果:双酶切实验证实得到pcDNA3-Flag-DEK阳性克隆;Western印迹实验发现DEK在293T细胞内表达;转录活性实验表明在ZR75-1乳腺癌细胞中,DEK呈剂量依赖性抑制p53启动子的活性。结论:构建了DEK的真核表达载体,并发现此表达载体能在ZR75-1乳腺癌细胞中抑制p53启动子活性。
Objective： To construct a eukaryotic expression vector for expression of DEK, and to study its effect on p53 promoter activity. Methods： The coding sequences of full length DEK was amplified from breast library by PCR and cloned into the pcDNA3-Flag vector. DEK expression was detected by Western blot after the recombi- nant plasmids were transfected into 293T cells. The activity of p53 promoter was detected in ZR75-1 cells. Re- sults： The full length DEK was expressed in the 293T cells. Compared to the empty vector, the activity of p53 promoter was decreased by DEK in a dose-dependent manner. Conclusion： We have constructed the eukaryotic ex- pression vector of DEK and found that p53 promoter activity was decreased by DEK in ZR75-1 cells.
出处
《生物技术通讯》
CAS
2013年第1期34-36,共3页
Letters in Biotechnology
基金
国家自然科学基金(31071174)
