p53 cDNA的克隆及在CV-1细胞中的表达

Eukaryotic Clone and Expression of p53 Gene

构建了可在哺乳动物细胞表达人p53基因的重组质粒pSV2neo-p53,并通过脂质体lipofectin介导转染猴肾细胞CV-1,用新霉素类抗生素G418筛选出阳性克隆。用原位杂交法检测转化细胞中的p53 mRNA;用免疫组化ABC法证明转化细胞的胞核中有p53蛋白表达。

It has been demonstrated that wild type p53 gene is a tumorsuppressor gene. Many human cancers are associated with the p53 mutations or the chromosome delections. It was shown that introduction of p53 gene into tumor cells in vitro can suppress the neoplastic phenotype and the cells growth rate. In this study, we constructed a human p53 reconbinant plasmid pSV2neo-p53. Lipofectin was employed to introduce DNA into normal monkey kidney cells w-1. G418 was used to select the dominant clones. It was shown that the p53 protein was expressed in the nuclei of transfected cells by adopting the immunohisto-chemistry ABC technique.