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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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基于发卡探针DNA/G-四链体模拟酶结构转化的高灵敏无标记荧光检测DNA
DOI: 10.11895/j.issn.0253-3820.150328, PP. 1402-1407
Keywords: 荧光,DNA,发卡探针,G-四链体模拟酶,结构转化
Abstract:
建立了基于目标DNA诱导的发卡探针DNA转化为G-四链体DNA过氧化物模拟酶的非标记荧光增强型DNA检测新体系。发卡探针DNA即分子信标探针由两部分构成,环状部分与目标DNA互补,茎部一端由一条富G序列构成。无目标DNA存在时,分子信标处于闭合状态,形成发卡结构;富G序列由于部分处于杂交状态,无法形成G四链体。当目标DNA与发卡探针DNA杂交并打开发卡结构,富G序列解链并自发折叠成G-四链体结构。G-四链体结构与血红素结合形成DNA模拟酶,催化H2O2还原的同时将无荧光的底物10-乙酰基-3,7-二羟基吩恶嗪(ADHP)氧化成荧光产物。通过测量荧光信号,实现对目标DNA的定量检测。优化后的体系检测条件为pH8.0,10mmol/LK+,0.2μmol/LHemin,50μmol/LADHP。在优化的条件下,目标DNA在0.005~1.0nmol/L浓度范围内与体系荧光信号呈线性关系,检出限为3.0pmol/L。本方法可以区分完全互补和单碱基错配的目标DNA。
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