【摘要】 为了使真核基因在真核细胞进行表达,我们将携带人白细胞介素4基因的表达质粒pdBHhIL-4转化哺乳动物细胞小鼠的C127细胞,该表达质粒由原始质粒pdKCR和牛乳头瘤病毒(pdBPV-1)及人IL-4 cDNA构建而成,再经酶切和凝胶电泳分析加以证实。细胞转化采用改进的磷酸钙沉淀法和甘油休克的方法,转化后的细胞逐渐呈现形态学上的改变,经活性测定其培养上清中具有人IL-4的表达。更多还原

【Abstract】 In order to express eukaryotic gene in eukaryotic eells, the expression plasmid pdBHhil-4 bring human interleukin 4 gene transformed into mammalian cells (mouse C127).The expression plasmid was constructed with partial DNA squences of plasmid pdKCR and bovine papilloma virus (pdBPV-1 ) as well as human IL-4 cDNA, which contains three bands of 5.12, 3.10 and 2.35 kb by analysis using restriction enzyme and gel electrophoresis.The transformation was performed with the calcium phosphate coprecipitation and glycerol shock, the transformed cloning cells exhibit altered cell morphology, loss of growth control and resulting in a dominant selective marker. The biological activity of IL-4 in culture supernatant of cloning C127 cells was identified ( 20 000IU/ml) .更多还原