卵巢癌的p53基因治疗实验研究——人野生型p53基因真核细胞表达质粒的构建

The study of p53 gene therapy for ovarian cancer ——Construction of recombinant mammalian expressing vector for human wild type p53 gene

目的 :本实验构建了人野生型 p53基因与真核表达载体 pc DNA3的重组体 ,为进行卵巢癌基因治疗研究打下基础。方法 :利用分子生物学基因克隆技术从质粒 p GEX-p53中用双酶切下目的片段 ,挤压法回收与 pc DNA3构成重组体。结果 :成功的构建了人野生型 p53基因真核细胞表达质粒。结论 :带有两个不同的酶切位点的目的片段定向克隆的方法使重组体的构建高效简便 ,挤压法回收目的片段经济有效。

Objective:To study the gene therapy for ovarian cancer,we constructed a human wild type p53 gene expressing vector in ovarian cancer cells.Methods:p53cDNA fragments were obtained by complete digestion of pGEX -p53 with BamHⅠ and EcoRⅠ.The fragments were purified by squeezing method and were inserted into the pcDNA3 to form an expressing vector.Results:We have constructed the recombinant mammalian expressing vector for human wild type p53 gene.Conclusions:Constructing a vector using the fragment with two cutting sites is efficient and simple.The squeezing method of purifying DNA is economic and efficient. 〔