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A Highly Sensitive and Selective Competition Assay for the Detection of Cysteine Using Mercury-Specific DNA, Hg2+ and Sybr Green I

机译:使用汞特异性DNA,Hg2 +和Sybr Green I检测半胱氨酸的高灵敏度和选择性竞争测定法

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摘要

We here report a rapid, sensitive, selective and label-free fluorescence detection method for cysteine (Cys). The conformation of mercury-specific DNA (MSD) changes from a random coil form to a hairpin structure in the presence of Hg2+ due to the formation of a thymine-Hg2+-thymine (T-Hg2+-T) complex. Cys can selectively coordinate with Hg2+ and extract it from the thymine-Hg2+-thymine complex. The hairpin structure dehybridizes and the fluorescence intensity of Sybr Green I (SG) decreases upon addition of Cys because SG efficiently discriminates mercury-specific DNA and mercury-specific DNA/Hg2+ complex. The detection can be finished within 5 min with high sensitivity and selectivity. In addition, we can obtain variable dynamic ranges for Cys by changing the concentration of MSD/Hg2+.
机译:我们在这里报告了一种快速,灵敏,选择性和无标记的半胱氨酸(Cys)荧光检测方法。由于胸腺嘧啶-Hg2 +-胸腺嘧啶(T-Hg2 + -T)复合物的形成,在Hg2 +存在下,汞特异性DNA(MSD)的构象从无规卷曲形式变为发夹结构。 Cys可以选择性地与Hg2 +配合并从胸腺嘧啶-Hg2 +-胸腺嘧啶复合物中提取。发夹结构去杂化并且Sybr Green I(SG)的荧光强度在添加Cys后降低,因为SG可有效区分汞特异性DNA和汞特异性DNA / Hg2 +复合物。检测可以在5分钟内以高灵敏度和选择性完成。此外,我们可以通过更改MSD / Hg2 +的浓度来获得可变的Cys动态范围。

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