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用大肠杆菌提取物对经X射线照射的脱氧核糖核酸进行核酸内切切割

Endonucleolytic incision of x-irradiated deoxyribonucleic acid by extracts of Escherichia coli.

作者信息

Strniste G F, Wallace S S

出版信息

Proc Natl Acad Sci U S A. 1975 Jun;72(6):1997-2001. doi: 10.1073/pnas.72.6.1997.

Abstract

An enconuclease activity that reacts with x-irradiated DNA is present in extracts of E. coli. By using centrifugal methods to monitor the conversion of the supercoiled, circular double-stranded DNA for phage phi-x-174 (replicative form) or PM2 to the relaxed circular form it was possible to quantitate the rate of radiation induced endonuclease-sensitive sites in the DNA. For every single-strand break induced by x-rays under aerobic irradiation conditions, there is approximately one induced site sensitive to this endonuclease activity. Under irradiation conditions (addition OF Potassium iodide) that dramatically reduce rates of single-strand breaks and "alkalilabile" lesions, the number of endonuclease-sensitive sites relative to single-strand breaks increase approximatley 4-fold. This nuclease is present in several strains of E. coli B and K12, including mutants deficient in DNA polymerase I, recombination gene products (rec mutants), ultraviolet light incision enzyme (uvr A mutant), and endonuclease II. It is suggested that this endonuclease may be involved in an excision repair process for damages incurred in DNA by ionizing radiation.

摘要

大肠杆菌提取物中存在一种能与经X射线照射的DNA发生反应的核酸内切酶活性。通过使用离心方法监测噬菌体φX174(复制型)或PM2的超螺旋环状双链DNA向松弛环状形式的转化,可以对DNA中辐射诱导的核酸内切酶敏感位点的速率进行定量。在有氧照射条件下,每一个由X射线诱导的单链断裂,大约有一个对这种核酸内切酶活性敏感的诱导位点。在显著降低单链断裂速率和“碱不稳定”损伤的照射条件下(添加碘化钾),相对于单链断裂,核酸内切酶敏感位点的数量增加约4倍。这种核酸酶存在于几种大肠杆菌B和K12菌株中,包括缺乏DNA聚合酶I、重组基因产物(rec突变体)、紫外线切割酶(uvr A突变体)和核酸内切酶II的突变体。有人提出,这种核酸内切酶可能参与了电离辐射对DNA造成损伤的切除修复过程。

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