Collins A, Jones C, Waldren C
J Cell Sci. 1982 Aug;56:423-40. doi: 10.1242/jcs.56.1.423.
We describe a rapid and simple assay for DNA damage and repair in mammalian cells, based on the partial unwinding of nicked DNA in alkali, and involving transfer of this DNA to nitrocellulose and the digestion of single-stranded DNA with S1 nuclease, all steps taking place on tissue-culture chamber slides. Direct breakage of DNA by ionizing radiation has been examined, and we have developed a standard procedure for measuring enzymic breakage of DNA as an index of excision-repair capacity following ultraviolet irradiation. We report a wide range of repair capacities among various hamster and human cell lines, with considerable overlap between the two species. Hybrids between hamster and human cells tend to display repair activity characteristic of the hamster parent.
我们描述了一种用于检测哺乳动物细胞中DNA损伤与修复的快速简便方法，该方法基于碱性条件下切口DNA的部分解旋，包括将此DNA转移至硝酸纤维素膜以及用S1核酸酶消化单链DNA，所有步骤均在组织培养室载玻片上进行。我们已研究了电离辐射对DNA的直接断裂作用，并且开发出一种标准程序，用于测量紫外线照射后DNA的酶促断裂，以此作为切除修复能力的指标。我们报告了多种仓鼠和人类细胞系之间广泛的修复能力范围，这两个物种之间存在相当大的重叠。仓鼠和人类细胞的杂种倾向于表现出仓鼠亲本特有的修复活性。
