Evaluation of DNA Damage Using Microwave Dielectric Absorption Spectroscopy
Evaluation of DNA Damage Using Microwave Dielectric Absorption Spectroscopy
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DOI：
10.14407/jrpr.2016.41.4.339
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发表时间：
2016-01-01
影响因子：
0.5
通讯作者：
Izumi, Yoshinobu
中科院分区：
文献类型：
--
作者：
Hirayama, Makoto;Matuo, Youichirou;Izumi, Yoshinobu
背景：评价脱氧核糖核酸（DNA）链断裂对于阐明电离辐射的生物效应具有重要意义。DNA-链断裂评价的常规方法已经通过琼脂糖凝胶电泳和其他使用DNA的电性质的方法实现。这种DNA链断裂评价系统可以根据DNA的分子量来评价DNA链断裂。然而，传统的方法需要样品的预处理和相对较长的分析周期。材料与方法：样品为水、甲醇和质粒DNA溶液。用大肠杆菌JM 109感受态细胞扩增质粒DNA pUC 118。用微波介电吸收光谱法测量了样品的谐振频率和Q值。当样品位于电场的中心时，作为驻波存在的频率的谐振曲线被干扰。结果与讨论：作为生物材料模型的甲醇浓度增加，共振频率向高频移动，Q值降低。双链断裂质粒DNA的微波功率谱的吸收峰从未损伤的质粒DNA偏移。此外，吸收峰的尖锐度发生变化，导致Q值发生变化。我们证实，共振频率移向更高的频率与质粒DNA.Conclusion浓度的增加：我们开发了一种新的技术用于评估DNA损伤。本文报道了利用微波介电吸收光谱技术评价DNA损伤的方法。
Background: Evaluation of deoxyribonucleic acid (DNA)-strand break is important to elucidate the biological effect of ionizing radiations. The conventional methods for DNA-strand break evaluation have been achieved by Agarose gel electrophoresis and others using an electrical property of DNAs. Such kinds of DNA-strand break evaluation systems can estimate DNAstrand break, according to a molecular weight of DNAs. However, the conventional method needs pre-treatment of the sample and a relatively long period for analysis. They do not have enough sensitivity to detect the strand break products in the low-dose region.Materials and Methods: The sample is water, methanol and plasmid DNA solution. The plasmid DNA pUC118 was multiplied by using Escherichia coli JM109 competent cells. The resonance frequency and Q-value were measured by means of microwave dielectric absorption spectroscopy. When a sample is located at a center of the electric field, resonance curve of the frequency that existed as a standing wave is disturbed. As a result, the perturbation effect to perform a resonance with different frequency is adopted.Results and discussion: The resonance frequency shifted to higher frequency with an increase in a concentration of methanol as the model of the biological material, and the Q-value decreased. The absorption peak in microwave power spectrum of the double-strand break plasmid DNA shifted from the non-damaged plasmid DNA. Moreover, the sharpness of absorption peak changed resulting in change in Q-value. We confirmed that a resonance frequency shifted to higher frequency with an increase in concentration of the plasmid DNA.Conclusion: We developed a new technique for an evaluation of DNA damage. In this paper, we report the evaluation method of DNA damage using microwave dielectric absorption spectroscopy.