Brash D E, Franklin W A, Sancar G B, Sancar A, Haseltine W A
J Biol Chem. 1985 Sep 25;260(21):11438-41.
The effect of purified Escherichia coli DNA photolyase on the UV light-induced pyrimidine-pyrimidone (6-4) photoproduct and cyclobutane pyrimidine dimer was investigated in vitro using enzyme purified from cells carrying the cloned phr gene (map position, 15.7 min). Photoproducts were examined both as site-specific lesions in end-labeled DNA and as chromatographically identified products in uniformly labeled DNA. E. coli DNA photolyase removed cyclobutane dimers but had no activity on pyrimidine-pyrimidone (6-4) photoproducts. Photoreactivation can therefore be used to separate the biological effects of these two UV light-induced molecular lesions.
使用从携带克隆的phr基因(图谱位置,15.7分钟)的细胞中纯化的酶,在体外研究了纯化的大肠杆菌DNA光解酶对紫外线诱导的嘧啶 - 嘧啶酮(6-4)光产物和环丁烷嘧啶二聚体的作用。光产物既作为末端标记DNA中的位点特异性损伤进行检测,也作为均匀标记DNA中色谱鉴定的产物进行检测。大肠杆菌DNA光解酶可去除环丁烷二聚体,但对嘧啶 - 嘧啶酮(6-4)光产物没有活性。因此,光复活作用可用于区分这两种紫外线诱导的分子损伤的生物学效应。