培养的人类细胞DNA中电离辐射诱导的损伤。8,5-环-2'-脱氧鸟苷的鉴定。
Ionizing-radiation-induced damage in the DNA of cultured human cells. Identification of 8,5-cyclo-2-deoxyguanosine.
作者信息
Dizdaroglu M, Dirksen M L, Jiang H X, Robbins J H
出版信息
Biochem J. 1987 Feb 1;241(3):929-32. doi: 10.1042/bj2410929.
Abstract
Epstein-Barr-virus-transformed peripheral-blood B-lymphocytes were gamma-irradiated at 0 degree C at doses from 10 to 100 Gy. The cells were immediately lysed and the DNA was isolated. Subsequently, the DNA was hydrolysed to 2'-deoxyribonucleosides with a mixture of DNAase I, venom and spleen exonucleases and alkaline phosphatase. The hydrolysate was dried, trimethylsilylated and analysed by capillary gas chromatography-mass spectrometry with selected-ion monitoring. The (5'R)- and (5'S)-diastereomers of 8,5'-cyclo-2'-deoxyguanosine were observed in a ratio of 1:3, and their formation was dose-dependent. It was possible to detect and characterize one such lesion in approx. 4 X 10(4) guanine nucleotide subunits of DNA.
摘要
将爱泼斯坦-巴尔病毒转化的外周血B淋巴细胞在0摄氏度下用10至100戈瑞的剂量进行γ射线照射。细胞立即裂解并分离出DNA。随后，用脱氧核糖核酸酶I、蛇毒和脾外切核酸酶以及碱性磷酸酶的混合物将DNA水解为2'-脱氧核糖核苷。水解产物干燥后进行三甲基硅烷化处理，并通过选择离子监测的毛细管气相色谱-质谱联用仪进行分析。观察到8,5'-环-2'-脱氧鸟苷的(5'R)-和(5'S)-非对映异构体的比例为1:3，且它们的形成呈剂量依赖性。在大约4×10(4)个DNA鸟嘌呤核苷酸亚基中能够检测并鉴定出一种这样的损伤。
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