摘 要:本文基于双链DNA为模板合成铜纳米颗粒具有荧光特性的原理,并以此做信号输出实现对DNA聚合酶活性的非标荧光检测。DNA发夹引物自身不能合成铜纳米颗粒,荧光较弱。目标DNA聚合酶诱导发夹引物的聚合延伸形成双链DNA产物,并以此为模板合成铜纳米颗粒,荧光显著增强。荧光强度与DNA聚合酶的浓度呈正比,检测荧光强度的变化实现对DNA聚合酶活性的检测。该方法原理简单,可操作性强且无需荧光基团标记,可推广适用于其他核酸修饰酶的检测。28281 毕业论文关键词:DNA聚合酶;铜纳米颗粒;DNA发夹引物;荧光检测
Double-Strand DNA-Templated Formation of Copper Nanoparticles as Fluorescent Probe for Label-free Detection of DNA Polymerase
Abstract:In this paper,we construct a label-free fluorescent assay for DNA polymerase based on the principle of double-strand DNA-templated formation of copper nanoparticles (Cu NPs),which can give fluorescence emission and be taken as signal readout.DNA hairpin primer (HP) cannot be taken as template for the formation of Cu NPs,and thus obtain a weak fluorescence.DNA polymerase can trigger the polymerization extension of HP and produce a double-stranded DNA (dsDNA),which then can be taken as the template for the formation of Cu NPs,and give a strong fluorescence signal.Fluorescence intensity is proportional to the concentration of DNA polymerase,and the detection of DNA polymerase can be realized by monitoring the changes of fluorescence intensity.The proposed method is simple,easy to operate and label free,it can be applied in the detection of other DNA modified enzymes.
Key words:DNA polymerase;Copper nanoparticles;DNA hairpin primer;Fluorescence detection
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