McCready S, Cox B
Department of Plant Sciences, University of Oxford, UK.
Mutat Res. 1993 Mar;293(3):233-40. doi: 10.1016/0921-8777(93)90074-q.
We have developed a dot blot immunoassay for UV photoproducts which distinguishes 6-4 photoproducts from cyclobutane dimers. The assay uses a polyclonal antiserum that is specific for UV-irradiated DNA. Cyclobutane dimers are measured in DNA samples which have been treated with hot alkali to destroy 6-4 photoproducts. 6-4 Photoproducts are measured using blots that have been incubated in photoreactivating enzyme to eliminate cyclobutane dimers. A combination of the two treatments leaves no detectable antigenic lesions. Wild-type S. cerevisiae repairs 6-4 photoproducts, in the genome overall, more rapidly than cyclobutane dimers. The most sensitive alleles of rad1, rad2, rad3 and rad4 are completely unable to repair either kind of photoproduct. We conclude that 6-4 photoproducts are repaired by essentially the same mechanism as are cyclobutane dimers.
我们开发了一种用于紫外线光产物的斑点印迹免疫测定法，该方法可区分6-4光产物和环丁烷二聚体。该测定法使用对紫外线照射的DNA具有特异性的多克隆抗血清。在已用热碱处理以破坏6-4光产物的DNA样品中测量环丁烷二聚体。使用在光复活酶中孵育以消除环丁烷二聚体的印迹来测量6-4光产物。两种处理的组合不会留下可检测到的抗原性损伤。野生型酿酒酵母在整个基因组中修复6-4光产物的速度比环丁烷二聚体更快。rad1、rad2、rad3和rad4最敏感的等位基因完全无法修复任何一种光产物。我们得出结论，6-4光产物的修复机制与环丁烷二聚体基本相同。
