中文摘要：

目的构建p53RFP表达载体,观察p53RFP表达上调对HEK293A细胞生长的影响。方法利用DNA克隆技术,将人p 53RFP基因的编码序列克隆至质粒pcDNA4/Myc-HisA,经限制性酶切、DNA测序鉴定重组载体。质粒扩增提取,经脂质体介导法转染HEK293A细胞,实时定量RT-PCR、Western blot检测转染后p53RFP mRNA及蛋白的表达水平。采用CCK-8检测p53RFP表达上调对HEK293A细胞增殖的影响。结果 p 53RFP转染后其mRNA及蛋白表达水平均显著上调;p 53RFP转染后72 h、96 h,细胞增殖受到显著抑制（P〈0.01）。结论成功构建p53RFP表达载体pcDNA4-p53RFP,并可在HEK293A细胞内有效表达,其表达可抑制HEK293A细胞增殖。

英文摘要：

Objective To construct a p53RFP expression vector and explore the effect of high expression of p53RFP on the proliferation of HEK293A cells. Methods The entire coding sequence of human p53RFP gene was cloned into pcDNA4/Myc-HisA vector by DNA cloning technique. The product was confirmed by enzymatic digestion and sequencing. The vector was amplified and extracted, then transfected into HEK293A cells via liposome, and the levels of p53RFP mRNA and protein were detected by quantitative real-time RT-PCR and Western blot. Cell proliferation of HEK293A was evaluated by CCK-8. Results The mRNA and protein levels in p53RFP both remarkably increased after HEK293A cells were transfected with p53RFP expression vector. Cell proliferation of HEK293A was significantly inhibited at the time points of 72 h and 96 h after p53RFP transfection （P 〈 0.01 ）. Conclusion The p53RFP expression vector was successfully constructed,which can be effectively transfected into HEK293A cells. The high expression of p53RFP could inhibit the cell proliferation in HEK293A.