A quantitative assay of deoxyribonucleic acid strand breaks and their repair in mammalian cells.

A quantitative assay of deoxyribonucleic acid strand breaks and their repair in mammalian cells.
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哺乳动物细胞中脱氧核糖核酸链断裂及其修复的定量测定。

DOI:
10.1016/0003-2697(82)90229-9
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发表时间:
1982
影响因子:
2.9
通讯作者:
Joner,EI
Joner,EI
中科院分区:
生物学4区
文献类型:
--
作者:
Blakely,WF;Ward,JF;Joner,EI

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碱性/滤膜DNA洗脱技术基于2 μ m孔径滤膜上大分子的DNA分子量依赖性保留,测量哺乳动物细胞中的单链DNA断裂。本文介绍了一种改进的技术,它使用[3 H]胸腺嘧啶标记的DNA的γ辐射的细胞作为内部参考。因此,在生物学上显著的辐射剂量(即,细胞存活发生的地方)。DNA损伤的测量基于实际的初始DNA洗脱速率,即,洗脱“测试”DNA的算术比率(即,14 C标记的DNA)相对于“参考”DNA(即,3 H标记的DNA)。在辐射后孵育的细胞中,这种损伤的修复被检测为“测试”DNA相对于来自未孵育细胞的“参考”DNA的凝集率的降低。对于用5戈伊(500拉德)照射的中国仓鼠V79-171细胞,修复可以分解为两个一级过程,其速率常数(在24°C下)分别为0.190和0.017 min-1。
The alkaline/filter DNA elution technique measures single-strand DNA breaks in mammalian cells based on the DNA molecular weight dependent retention of the macromolecule on 2-μm-pore-size filters. Described here is a modification of the technique which uses [3H]thymidine-labeled DNA of γ-irradiated cells as an internal reference. Thus, an increased precision is obtained in the assessment of this type of DNA damage at biologically significant radiation doses (i.e., where cell survival occurs). The measure of DNA damage is based on the actual initial DNA elution rate, i.e., arithmetic ratio of the elution of “test” DNA (i.e.,14C-labeled DNA) relative to the elution of “reference” DNA (i.e.,3H-labeled DNA). The repair of this damage on postirradiation incubation of the cells is detected as a decrease in the rate of “test” DNA cluted relative to “reference” DNA from unincubated cells. For Chinese hamster V79–171 cells irradiated with 5 Gy (500 rads), repair can be resolved into two first-order processes having rate constants (at 24°C) of ∼0.190 and ∼0.017 min−1.
使用非放射性碱性洗脱技术评估体内诱导的 DNA 损伤。
DOI: 10.1016/s0304-3835(79)80037-3
发表时间: 1979
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影响因子: 3.1
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DOI: --
发表时间: 1973
期刊: International Journal of Radiation Biology and Related Studies in Physics Chemistry and Medicine
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检测人类细胞中诱导的 DNA 链断裂,灵敏度更高。
DOI: --
发表时间: 1980
期刊: Radiation Research
影响因子: 3.4
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DOI: --
发表时间: 1967
期刊: Nature
影响因子: 64.8
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