Hunting D J, Dresler S L, de Murcia G
Biochemistry. 1985 Oct 8;24(21):5729-34. doi: 10.1021/bi00342a007.
Biotin-labeled deoxyuridine triphosphate (BiodUTP) has the potential to be a useful affinity probe for studies on DNA repair, if it can be incorporated into DNA repair patches and does not inhibit subsequent steps in the excision repair pathway. We have synthesized BiodUTP by an improved procedure and have used permeable normal human fibroblasts to determine the effect of substituting BiodUTP for thymidine triphosphate on several steps in the excision repair pathway: incision, polymerization, ligation, and nucleosome rearrangement. The results demonstrate that BiodUTP is efficiently incorporated into repair patches and has little or no effect on the repair process. The presence of BiodUMP in ligated repair patches has been used to visualize the repair patches by electron microscopy following incubation with ferritin-labeled avidin. This approach has been used to estimate the maximum size of repair patches induced by ultraviolet radiation.
生物素标记的三磷酸脱氧尿苷（BiodUTP）如果能够掺入DNA修复片段且不抑制切除修复途径中的后续步骤，就有可能成为研究DNA修复的有用亲和探针。我们通过改进的方法合成了BiodUTP，并使用可渗透的正常人成纤维细胞来确定用BiodUTP替代三磷酸胸苷对切除修复途径中几个步骤的影响：切口、聚合、连接和核小体重排。结果表明，BiodUTP能有效地掺入修复片段，对修复过程几乎没有影响。在与铁蛋白标记的抗生物素蛋白孵育后，通过电子显微镜观察连接的修复片段中生物素化单磷酸脱氧尿苷（BiodUMP）的存在情况来可视化修复片段。这种方法已被用于估计紫外线辐射诱导的修复片段的最大尺寸。
