携带p53基因重组腺病毒载体的构建

Construction of p53 Recombinant Adenovirus Vector

将 pCMVp5 3重组转移载体经BamHI和NheI酶切 ,得到 p5 3基因cDNA ,然后将cDNA片段克隆到转移载体 pCMV5GFP ,使其受CMV5启动子的调控 ,获得 pCMV5p5 3重组转移载体 .用该线状重组转移载体与腺病毒右臂DNA经磷酸钙共转染 2 93细胞获得重组腺病毒 ,经酶联免疫吸附法 (ELISA)测定 p5 3蛋白含量 ,证明外源 p5 3基因在含重组腺病毒的 2

p53 cDNA is cloned into shuttle vector pCMV5GFP to get p53 recombinant adenovirus vector and make it controlled by CMV5 promotor and enhancer. And p53 cDNA is rescued into adenovirus by calcium phosphate cotransfection of linearized pCMV5p53 and QBI viral DNA into 293 cells and positive recombinant adenovirus plaques are screened by fluores centmicroscope. Expression of p53 gene in 293 cell is detected by EALISA. p53 recombinant adenovirus vector can be constructed and used for further research.