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目的 构建源于PNP/Mep -dR系统的融合及单自杀基因表达载体并检测二者表达。方法 经DNA重组技术制备融合基因PNP -TK ,将其及PNP基因分别插入真核表达载体pcDNA3 0 ,构建两真核表达载体pcDNA3 0 /PNP -TK和pcDNA3 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表达载体并在HepG2细胞株中检测到二者表达。结论 两个重组体 ,尤其是pcDNA3 0 /PNP -TK ,是肿瘤基因治疗中高效、广谱的理想载体。
Objective To construct the fusion and single suicide gene expression vector derived from PNP / Mep-dR system and test the expression of the two genes. Methods The fusion gene PNP -TK was prepared by DNA recombination technique. The two fusion proteins were inserted into eukaryotic expression vector pcDNA3 0 to construct two eukaryotic expression vectors, pcDNA3 0 / PNP -TK and pcDNA3 0 / PNP. Identification of two recombinants. Results Two novel suicide gene expression vectors were successfully constructed and their expression was detected in HepG2 cell line. Conclusion The two recombinants, especially pcDNA3 0 / PNP-TK, are ideal carriers for efficient and broad-spectrum tumor gene therapy.