Visual quantification of DNA double-strand breaks in bacteria
Visual quantification of DNA double-strand breaks in bacteria
复制标题
DOI：
10.1016/s0027-5107(99)00124-4
复制
发表时间：
1999-10-19
影响因子：
2.3
通讯作者：
Lai, H
中科院分区：
文献类型：
--
作者：
Singh, NP;Stephens, RE;Lai, H
在这篇文章中，我们描述了一种显示单个电拉伸的大肠杆菌DNA分子中双链断裂的方法。我们还提供了在中性微凝胶电泳条件下电拉伸或迁移的DNA由单个染色体组成的证据。用中性微凝胶电泳法测定0、12.5、25、50和100rad X射线照射后的DNA迁移(拉伸)和DNA双链断裂数。强烈荧光染料YOYO和定制幻灯片的使用帮助我们可视化了单个细菌DNA分子。细菌DNA在结构上与人类淋巴细胞的电拉伸DNA相似。我们能够检测到低至12.5rad的X射线剂量引起的DNA迁移(拉伸)的变化，以及低至25rad的剂量引起的DNA断裂数量的增加。DNA迁移的程度和断裂数目与X射线剂量直接相关。(C)1999 Elsevier Science B.V.保留所有权利。
In this paper, we describe a method for the visualization of double-strand breaks in a single electrostretched Escherichia coli DNA molecule. We also provide evidence that electrostretched or migrated DNA under neutral microgel electrophoresis conditions is made up of individual chromosomes. Using the neutral microgel electrophoresis technique, DNA migration (stretching) was measured and the number of DNA double-strand breaks were counted following exposure of E. coli cells to 0, 12.5, 25, 50, or 100 rad of X-rays. The use of an intense fluorescent dye, YOYO and custom-made slides have helped us in visualizing individual bacterial DNA molecules. Bacterial DNA appears similar in structure compared to electrostretched DNA from human lymphocytes. We were able to detect changes in DNA migration (stretching) induced by an X-ray dose as low as 12.5 rad and an increase in the number of DNA breaks induced by a dose as low as 25 rad. The extent of DNA migration and number of breaks were directly correlated to X-ray dosage. (C) 1999 Elsevier Science B.V. All rights reserved.