Fluorescence Detection of Single‐Nucleotide Polymorphism with Single‐Strand Triplex‐Forming DNA Probes
Fluorescence Detection of Single‐Nucleotide Polymorphism with Single‐Strand Triplex‐Forming DNA Probes
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DOI：
10.1002/cbic.201100534
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发表时间：
2011-12
期刊：
影响因子：
3.2
通讯作者：
Xinpeng Li;Y. Wang;Jiajie Guo;Xinjing Tang
中科院分区：
文献类型：
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作者：
Xinpeng Li;Y. Wang;Jiajie Guo;Xinjing Tang
三螺旋形成寡核苷酸 (TFO) 在基因组中广泛存在，并且已在调控区域中发现，特别是在 DNA 的启动子区和重组热点中。为了特异性检测这些多嘌呤序列，我们设计并合成了两种双芘标记的单链寡核苷酸探针（TFO-FP），由识别序列、连接序列和检测序列组成。 TFO-FP 与目标多嘌呤寡核苷酸的杂交过程涉及 Watson-Crick 和 Hoogsteen 碱基配对。通过寡核苷酸序列的双重传感，通过监测芘荧光的变化来检测目标寡核苷酸的单突变。探针的高特异性在很宽的温度范围内保持，而不牺牲杂交动力学。
Triple‐helix‐forming oligonucleotides (TFOs) are widespread in the genome and have been found in regulatory regions, especially in promoter zones and recombination hotspots of DNA. To specifically detect these polypurine sequences, we designed and synthesized two dual pyrene‐labeled single‐strand oligonucleotide probes (TFO‐FPs) consisting of recognition, linker, and detection sequences. The hybridization processes of TFO‐FPs with target polypurine oligonucleotides involve both Watson–Crick and Hoogsteen base‐pairings. Through double sensing of oligonucleotide sequences, single mutations of target oligonucleotides are detected by monitoring changes in pyrene fluorescence. The high specificities of the probes are maintained over a wide temperature range without sacrifice of hybridization kinetics.