人野生型p53基因与逆转录病毒载体的构建及转染  

CONSTRUCTION AND TRANSFECTION OF PLSAMID OF HUMAN WILD TYPE p53 GENE IN RETROVIRAL VECTOR

作　　者：吴岩[1] 李文梅[1] 张青云[1] 吕有勇[1] 舍英[1] 

机构地区：[1]北京市肿瘤防治研究所,内蒙古医学院组胚教研室

出　　处：《内蒙古医学院学报》1995年第1期11-15,共5页Acta Academiae Medicinae Neimongol

基　　金：国家863攻关课题

摘　　要：应用逆转录病毒为载体构建了含人野生型ｐ５３基因的重组质粒，并成功地转染ψｃｒｉｐ包装细胞。本实验应用ＰＬＸＳＮ为载体，１．８ｋｂ野生型ｐ５３基因作为插入片段，通过Ｔ＿４ＤＮＡ连接酶进行连接。用〔α—￣（３２）Ｐ〕ｄＣＴＰ标记野生型ｐ５３基因片段（１．８ｋｂ）为探针进行原位杂交。筛选出重组体，将其命名为ＰＬＸＳＮ－５３，然后用该重组体对ψｃｒｉｐ包装细胞进行转染。收集转染后的细胞，准备下一步感染ｐ５３突变细胞株研究之用。该质粒的构建和细胞转染实验为进一步研究野生型ｐ５３基因的抑癌作用及其表达与调控原理奠定了基础。Retrovirl evctor was used to construct a plasmid containing human wild type p53 gene and theplasmid was transfected to ψcirp packaging cells.PLXSN as a vector and a 1.8 kb fragment of wild type p53as a target gene were used in this study,and ligated with T_4DNA ligase. Positive clones being recombinantswere selected with ￣(32)p-labeled 1. 8kb fragment of wild type p53 as a probe.We named the recombinantsPLXSN-53,then tranfected them toψcirp packaging cells and. after collecting the surviving clones,transfected them to p53 mutation cell lines.The construction of PLXSN-53 and the success in trasfectingthem to ψcrip have provided a foundation for studying inhibition of tumor by xildd type p53 as well asmechehism of resulation,and expression of p53.

关 键 词：肿瘤 肿瘤抑制基因 逆转录病毒 载体 转染 

分 类 号：R730.21[医药卫生—肿瘤]