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两种基于PNP/ Mep-dR 系统的新型融合自杀基因载体的构建及鉴定

Constructions and Identif ication of two Fusion Gene Vectors Based on PNP/ Mep-dR Suicide Gene System

  • 摘要: 目的 构建两个新型融合自杀基因表达载体并检测其表达。方法 利用DNA重组技术制备两个融合基因PNP-TK和PNP-CD,将二者分别插入真核表达载体pcDNA3.0中,构建两个融合基因表达载体pcDNA3.0/PNP-TK和pcDNA3.0/PNP-CD;经酶切、PCR及测序鉴定各个重组体。结果 成功构建了两个新型融合基因表达载体并在肝癌细胞株HepG2中检测到二者的表达。结论 两个基于PNP/Mep-dR系统的融合自杀基因表达载体是肿瘤基因治疗中杀瘤谱广、杀伤效应强大的理想载体。

     

    Abstract: Objective  To const ruct two novel fusion suicide gene expression vectors, and detect their expression in human hepatocellular carcinoma cell line Hep G2. Methods  Two fusion suicide gene PNP-TK and PNP-CD were generated respectively by recombinant DNA techniques. Two fusion genes were inserted into pcDNA 3. 0 vector separately to const ruct two suicide gene expression vectors, pcDNA 3. 0/PNP-TK and pcDNA 3. 0/ PNP-CD. Two recombinants were analyzed and identified by recombinant enzyme, PCR and sequencing. Then they were t ransfected into human hepatocellular carcinoma cell line Hep G2 by liposome-mediated method. The expressions of two fusion genes were detected in Hep G2 cell ine. Results  Two fusion genes were const ructed successfully and the expression of two fusion genes were detected in human hepatocellular carcinoma cell line Hep G2. Conclusion  Two fusion suicide gene expression vectors based on PNP/ Mep-dR system are effective and broad-spect rum vectors for human hepatocellular carcinoma gene therapy.

     

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