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目的:构建含有tk基因的真核表达载体,并转染C6细胞株建立其表达系统,为探索胶质瘤的基因治疗奠定实验基础。方法:通过酶切pSNAV2.0-TK和pcDNA3分别获得目的基因tk片段及pcDNA3片段,将目的基因插入载体相应的酶切位点,构建pcDNA3-TK质粒。酶切鉴定后采用脂质体法瞬时转染C6细胞,RT-PCR检测tk基因在细胞中的表达。结果:pcDNA3-TK质粒构建成功并顺利导入C6细胞中。结论:构建的pcDNA3-TK质粒能够在转染的C6细胞中稳定、持续和高效地表达。
OBJECTIVE: To construct eukaryotic expression vector containing tk gene and transfect it into C6 cell line to establish its expression system, which laid the experimental foundation for exploring the gene therapy of glioma. Methods: The target gene tk fragment and pcDNA3 fragment were obtained by digesting pSNAV2.0-TK and pcDNA3 respectively. The target gene was inserted into the corresponding restriction sites of the vector to construct the pcDNA3-TK plasmid. After digestion and identification, the C6 cells were transiently transfected by lipofectamine and the expression of tk gene was detected by RT-PCR. Results: The pcDNA3-TK plasmid was successfully constructed and successfully transfected into C6 cells. CONCLUSION: The constructed pcDNA3-TK plasmid can stably, consistently and efficiently express in transfected C6 cells.