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目的 :制备具有治疗活性的野生型p5 3基因真核表达载体的细胞疫苗。方法 :利用RT PCR从人外周血淋巴细胞总mRNA中扩增出全长的野生型p5 3基因 ,将其克隆入TA克隆载体 ,经扩增后 ,将p5 3cDNA连入pCMV真核表达载体中。利用lipofectamine将其转染DU145细胞并通过免疫组化的方法检测其表达情况。结果 :p5 3基因可在DU145细胞中稳定表达。结论 :制备了可稳定表达于DU145细胞中的野生型p5 3基因细胞疫苗。
Objective : To prepare a cell vaccine with a therapeutically active wild type p53 gene eukaryotic expression vector. METHODS: The full-length wild-type p53 gene was amplified from the total mRNA of human peripheral blood lymphocytes by RT PCR and cloned into a TA cloning vector. After amplification, the p53 cDNA was ligated into the pCMV eukaryotic expression vector. . The lipofectamine was used to transfect DU145 cells and its expression was detected by immunohistochemistry. Results: The p53 gene was stably expressed in DU145 cells. Conclusion: A wild-type p53 gene cell vaccine that can be stably expressed in DU145 cells was prepared.