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Hg2+ detection using a phosphorothioate RNA probe adsorbed on graphene oxide and a comparison with thymine-rich DNA

机译:使用吸附在氧化石墨烯上的硫代磷酸RNA探针检测Hg2 +,并与富含胸腺嘧啶的DNA进行比较

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摘要

Mercury is a highly toxic heavy metal and many DNA-based biosensors have been recently developed for Hg2+ detection in water. Among them, thymine-rich DNA is the most commonly used for designing Hg2+ sensors. However, the thymine-Hg2+ interaction is strongly affected by the buffer conditions. We recently reported a molecular beacon containing phosphorothioate (PS)-modified RNA linkages that can be cleaved by Hg2+. In this work, the fluorescence quenching and DNA adsorption properties of nano-sized graphene oxide (NGO) were used to develop a new sensor using the PS-RNA chemistry. Three DNA probes, containing one, three and five PS-RNA linkages, respectively, were tested. Finally, a fluorophore-labeled poly-A DNA with five PS-RNA linkages was selected and adsorbed by NGO. In the presence of Hg2+, the fluorophore was released from NGO due to the cleavage reaction, resulting in a fluorescence enhancement. This sensor is highly selective for Hg2+ with a detection limit of 8.5 nM Hg2+. For comparison, a fluorophore-labeled poly-T DNA was also tested, which responded to Hg2+ more slowly and was inhibited by high NaCl concentrations, while the PS-RNA probe was more tolerant to different buffer conditions. This work indicates a new method for interfacing DNA with NGO for Hg2+ detection.
机译:汞是一种剧毒的重金属,最近开发了许多基于DNA的生物传感器来检测水中的Hg2 +。其中,富含胸腺嘧啶的DNA是设计Hg2 +传感器最常用的DNA。但是,胸腺嘧啶-Hg2 +的相互作用受缓冲液条件的强烈影响。我们最近报道了一个分子信标,其中包含可以被Hg2 +裂解的硫代磷酸酯(PS)修饰的RNA键。在这项工作中,纳米级氧化石墨烯(NGO)的荧光猝灭和DNA吸附特性被用于开发一种使用PS-RNA化学方法的新型传感器。测试了三个分别包含一个,三个和五个PS-RNA连接的DNA探针。最后,选择具有五个PS-RNA键的荧光团标记的poly-A DNA并用NGO吸附。在Hg2 +存在下,由于裂解反应,荧光团从NGO释放出来,导致荧光增强。该传感器对Hg2 +具有高度选择性,检测极限为8.5 nM Hg2 +。为了进行比较,还测试了荧光团标记的poly-T DNA,其对Hg2 +的反应较慢,并被高浓度的NaCl抑制,而PS-RNA探针对不同的缓冲液条件更耐受。这项工作表明了一种将DNA与NGO进行Hg2 +检测的新方法。

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