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An Endonuclease from Escherichia coli That Acts Preferentially on UV-Irradiated DNA and Is Absent from the uvrA and uvrB Mutants*

机译:来自大肠杆菌的核酸内切酶,主要作用于紫外线辐射的DNA,而在uvrA和uvrB突变体中不存在

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摘要

At least two endonucleolytic activities that preferentially incise ultraviolet (UV)-irradiated DNA exist in extracts of E. coli. These two activities can be separated by phosphocellulose chromatographic fractionation. The subject of this paper is one of these activities, which elutes from phosphocellulose with 0.25 M potassium phosphate, pH 7.5. This endonucleolytic activity specific for UV-irradiated DNA is absent from partially purified extracts of uvrA and uvrB mutants, which are defective in excision of pyrimidine dimers, but is present in normal amounts in the uvrC excision-defective mutant. The enzyme binds very tightly and specifically to UV-irradiated DNA. Binding can be prevented by prior treatment of the irradiated DNA with photoreactivating enzyme. This binding activity is absent in uvrA and uvrB mutants, but present in uvrC and uvrD mutants.
机译:大肠杆菌提取物中至少存在两种​​优先切割紫外线(UV)辐射的DNA的核酸内切酶活性。这两个活性可以通过磷酸纤维素色谱分离分离。本文的主题是这些活性之一,它是用0.25 M磷酸钾,pH 7.5从磷酸纤维素中洗脱出来的。 uvrA和uvrB突变体的部分纯化提取物不存在这种对UV辐射的DNA特有的内切核酸活性,这些提取物在嘧啶二聚体的切除中有缺陷,但在uvrC切除缺陷的突变体中以正常量存在。该酶与紫外线辐射的DNA紧密结合,特异性结合。可以通过用光活化酶预先处理被辐照的DNA来防止结合。该结合活性在uvrA和uvrB突变体中不存在,但在uvrC和uvrD突变体中存在。

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