重组P_(53)基因表达及其与Fas关系初探

Preliminarily Investigating the Expression of theRecombinant P53 Gene and the Relation between the P53 Gene Expression and Fas Gene Expression

为观察重组ＰｘＴ１－Ｐ５３在真核细胞中的表达及其与Ｆａｓ表达的关系，笔者采用磷酸钙—ＤＮＡ共沉淀法，ＤＥＡＥ—莆聚糖介导的ＤＮＡ转染法将ＰｘＴ１－Ｐ５３转移至ＰＡ３１７细胞内，并用抗Ｐ５３和抗Ｆａｓ抗体，经间接免疫荧光法证实转染ＰｘＴ１－Ｐ５３的ＰＡ３１７细胞不仅能表达Ｐ５３蛋白，而且亦能表达Ｆａｓ抗原。本实验结果表明笔者构建的ＰｘＴ１－Ｐ５３确实能在真核细胞内表达，并提示Ｐ５３可能通过其转录因子活性，诱导Ｆａｓ等基因表达。

おummary:In order to investigate if the constructed recombinant P53 gene could express in the eukaryotes and the relation between P53 gene and Fas gene expression,the 12.2kb recombinant P53 gene was transfirred into the mouse PA317 cell line by the calcium phosphate-DNA coprecipitation method and DEAE-Dextran transfection method,then,the anti-P53 and anti-Fas were used to analyse the expression of P53 and Fas gene in the PA317 cell by indirect Immunofluorecent labeling technique.The Obtained experimental results confirmed the PA317 cells could express the recombinant P53 and suggested the expressing P53 protein could induce Fas gene and others gene to express by the transcriptional activities of P53 proteir to mediated the apoptosis of the cells with damaged DNA.