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观察了受照射小鼠瓦尔代尔扁桃体环 (WRE)在体细胞出现DNA双链断裂修复的忠实性 ,用6 0 Coγ射线照射后观察WRE细胞凋亡的最高峰 ,在此时间点处死小鼠 ,取WRE细胞分别用脂质体和电穿孔方法介导的基因转染技术 ,将双标记基因质粒 pPMH16导入细胞。研究WRE细胞DNA双链修复的忠实性。结果表明 ,未照射WRE细胞的抗G4 18转化克隆有 6 9.75 %的 gpt基因表达 ,说明大部分断裂的 gpt基因被忠实性地修复 ;2Gy、4Gy射线照射的WRE细胞的转化克隆只有 36 .0 5 %和 2 1.5 0 %的克隆能正确表达 gpt基因功能 ,说明WRE细胞照射后DNA双链断裂的修复显著低于正常细胞 ,且剂量越大 ,修复的忠实性越低。脂质体和电穿孔方法介导的基因转染技术可用于原代细胞基因转染的研究 ,照射后的WRE细胞易发生DNA双链断裂错误重接。
The faithfulness of DNA double strand breaks in the somatic cells of irradiated mice was observed. The highest peak of WRE apoptosis was observed after irradiated with 60 Co γ ray. At this time point, the mice were sacrificed The double marker gene plasmid pPMH16 was transfected into the cells by using gene transfection technology mediated by liposome and electroporation method respectively. To study the faithfulness of DNA double-strand repair in WRE cells. The results showed that 6 .75% of the gpt gene was expressed in the anti-G4 18 transformants without irradiation of WRE cells, indicating that most of the cleaved gpt genes were faithfully repaired; the transformation clones of 2Gy and 4Gy-irradiated WRE cells were only 36.0 The 5% and 21.5% clones could correctly express the function of gpt gene, indicating that the repair of DNA double-strand breaks in WRE cells was significantly lower than that in normal cells. The larger the dose, the lower the loyalty of the repair. Liposomes and electroporation-mediated gene transfection technology can be used for primary cell gene transfection research, WRE cells after irradiation prone to DNA double-strand breaks fault reconnection.