Lohrer H D, Tangen U, Anderson R F, Arrand J E
Cancer Research Campaign Gray Laboratory, Mount Vernon Hospital, Northwood, Middlesex, UK.
Pathobiology. 1994;62(3):140-8. doi: 10.1159/000163894.
The fibroblast cell line AT5BIVA, from ataxia telangiectasia complementation group D, was transfected with a neo gene providing G418 resistance for subsequent selections. The G418 resistant cell line was fused with gamma-irradiated Chinese hamster ovary cells and a radioresistant hybrid, atxbc, was isolated following an X-ray selection procedure. All the cellular defects characteristic of ataxia telangiectasia were corrected to some degree; atxbc cells: (i) were confirmed to be resistant to X-rays; (ii) had regained control over DNA synthesis after DNA damage, and (iii) could overcome the radiation-induced block in the G2 phase of the cell cycle. Repetitive element polymerase chain reaction amplification of integrated hamster DNA from primers to middle repetitive elements confirmed the presence of hamster-specific sequences, suggesting that the phenotype of the cells had been corrected by integration of a normal hamster gene rather than by a reversion event.
来自共济失调毛细血管扩张症互补组D的成纤维细胞系AT5BIVA用提供G418抗性的新基因进行转染,用于后续筛选。将G418抗性细胞系与经γ射线照射的中国仓鼠卵巢细胞融合,并通过X射线选择程序分离出抗辐射杂种细胞atxbc。共济失调毛细血管扩张症的所有细胞缺陷都在一定程度上得到了纠正;atxbc细胞:(i)被证实对X射线有抗性;(ii)在DNA损伤后恢复了对DNA合成的控制,并且(iii)能够克服辐射诱导的细胞周期G2期阻滞。从针对中度重复元件的引物对整合的仓鼠DNA进行重复元件聚合酶链反应扩增,证实了仓鼠特异性序列的存在,表明细胞的表型是通过整合正常仓鼠基因而非回复事件得到纠正的。