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目的 观察腺相关病毒和人γ 干扰素的重组体 pWIG转染淋巴细胞及在细胞内表达情况。方法 将 pWIG以DNA 磷酸钙共沉淀法转染人淋巴细胞株H9,再分别用聚合酶链反应 (PCR)、逆转录 PCR(RT PCR)和酶联免疫吸附试验 (ELISA)等方法在DNA、RNA和蛋白质水平研究该重组体导入细胞和在细胞内表达的情况。结果 转染 pWIG的H9细胞的DNA和RNA经PCR和RT PCR检测均有 5 0 4bp的DNA条带 ;用ELISA可测出γ 干扰素的表达量为 17.2 pg/ 3× 10 6细胞。结论 重组体 pWIG用DNA 磷酸钙共沉淀法导入细胞后 ,转录HuIFN γmRNA ,并翻译人γ 干扰素蛋白。为基因治疗的进一步研究打下基础
Objective To observe the expression of pWIG-transfected lymphocytes and their expression in the cells of adeno-associated virus and human interferon-γ. METHODS: Human lymphocyte cell line H9 was transfected with pWIG by DNA-CaPc. The expression of pGEM was detected by polymerase chain reaction (PCR), reverse transcriptase PCR (RT PCR) and enzyme-linked immunosorbent assay (ELISA) RNA and Protein Levels The recombinant is introduced into cells and expressed intracellularly. Results The DNA and RNA of H9 cells transfected with pWIG were all detected by PCR and RT PCR. The band of 504 bp was detected. The expression of IFN-γ was 17.2 pg / 3 × 10 6 by ELISA. Conclusion Recombinant pWIG was transfected into cells with DNA-CaPc and transcribed HuIFNγmRNA and translated human interferon gamma protein. Lay the foundation for further research on gene therapy