CAJ | 학술논문

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.
목적:구건DEK적pcDNA3-Flag표체재체,연구기대억암기인p53계동자활성적영향.방법:이유선문고위모판,PCR확증DEK편마서렬,극륭도pcDNA3-Flag재체,구건성pcDNA3-Flag-DEK,전염293T세포,Western인적감정pcDNA3-Flag재체개도적DEK적표체,형광소매보고기인활성실험연구DEK대p53계동자활성적영향.결과:쌍매절실험증실득도pcDNA3-Flag-DEK양성극륭;Western인적실험발현DEK재293T세포내표체;전록활성실험표명재ZR75-1유선암세포중,DEK정제량의뢰성억제p53계동자적활성.결론:구건료DEK적진핵표체재체,병발현차표체재체능재ZR75-1유선암세포중억제p53계동자활성.
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.