一种用于检测产生自由基介导的DNA链断裂的试剂的菌株

B. Salles, M. Germanier, M. Defais
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引用次数: 12

摘要

在携带两个突变的大肠杆菌菌株中,一个在参与DNA复制起始的dnaC基因中,另一个在影响切除-修复系统的uvrB基因中,已经证明紫外线不能诱导SOS反应。这可能是由于缺乏任何诱导信号(Salles和Defais, 1984)。以RecA蛋白扩增为探针,观察其诱导SOS网络的能力。当DNA产生断裂时,RecA蛋白的诱导恢复。我们在这里描述了一种菌株,其中来自sfiA::lacZ融合的RecA蛋白和β-半乳糖苷酶可以在同一细菌提取物中同时测量。在没有复制进行的条件下,这种菌株可以用来检测化学物质在体内产生自由基介导的DNA断裂的能力。
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A bacterial strain for detecting agents that produce free radical-mediated DNA strand breaks

In an E. coli strain carrying two mutations, one in the dnaC gene involved in initiation of DNA replications and another in the uvrB gene which affects the excision-repair system, it has been shown that the SOS response cannot be induced by UV. This is probably due to the absence of any inducing signal (Salles and Defais, 1984). The capacity to induce the SOS network was followed using RecA protein amplification as a probe. When breaks were produced in DNA, RecA protein induction was restored. We describe here a strain in which both RecA protein and β-galactosidase from a sfiA::lacZ fusion can be measured simultaneously in the same bacterial extract. In conditions in which no replications proceeds, this strain can be used to detect the ability of chemicals to produce free radical-mediated DNA breaks in vivo.

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