【摘要】 目的 :构建新型PNP/Mep dR自杀基因系统的融合和单基因表达载体。方法 :利用DNA重组技术制备融合自杀基因PNP CD ,将其和PNP单基因分别插入真核表达载体pcDNA 3 0中 ,构建融合与单自杀基因表达载体pcDNA 3 0 /PNP CD和pcD NA 3 0 /PNP ;酶切、PCR及测序鉴定两重组体。结果 :成功构建了两个新型自杀基因载体并实现在HepG2肝癌细胞株中的表达。结论 :两个新型自杀基因表达载体 ,尤其是PNP CD融合自杀基因载体 ,是肿瘤基因治疗中广谱、高效的治疗载体。更多还原

【Abstract】 OBJECTIVE: To construct a fusion gene and a single gene expression vectors based on a novel PNP/Mep-dR suicide gene system. METHODS:The PNP-CD fusion suicide gene was generated by recombinant DNA techniques. The fusion gene and the single gene were inserted into pcDNA 3.0 vector separately to construct two suicide gene expression vectors, pcDNA 3.0/PNP-CD and pcDNA 3.0/PNP. Two recombinants were analyzed and identified by recombinant enzyme, PCR and sequencing. Then they were transfected into human hepatocellular carcinoma cell line HepG2 by the liposome-mediated method. The expressions of two genes were detected in cell line HepG2.RESULTS:The constructions of a fusion gene and a single gene expression vectors on the basis of the novel PNP/Mep-dR suicide gene system were successful and the expressions of PNP gene were detected in human hepatocellular carcinoma cell line HepG2. CONSLUSIONS:Two expression vectors based on the PNP/Mep-dR suicide gene system, especially the vector harboring the PNP-CD fusion gene, are effective and broad-spectrum vectors for the gene therapy of tumors.更多还原