Unambiguous detection of target DNAs by excimer-monomer switching molecular beacons
Unambiguous detection of target DNAs by excimer-monomer switching molecular beacons
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DOI：
10.1021/jo049824f
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发表时间：
2004-05-14
影响因子：
3.6
通讯作者：
Inouye, M
中科院分区：
文献类型：
--
作者：
Fujimoto, K;Shimizu, H;Inouye, M
开发了一类新型分子信标，其中芘荧光团连接在单链寡核苷酸的 3' 和 5' 端。两种基于芘的荧光团由相同的起始材料合成，从而简化了信标的制备。目标 DNA 信标的检测策略基于芘荧光团的“准分子-单体发射转换”：当信标与目标杂交时，芘部分的准分子发射变为单体 1。这种类型的荧光二态模式可以明确检测目标 DNA，因为可以通过荧光变化的等发射点的存在来监测非杂交和杂交信标之间严格的 1:1 相关性。信标可以检测目标 19 聚体 DNA，并可以在 1 nM 浓度下将目标与其单核苷酸错配区分开来。与传统分子信标相比，讨论了准分子-单体切换分子信标的优点。
A new class of molecular beacons were developed in which pyrene fluorophores were connected both at 3' and 5' ends of a single-stranded oligonucleotide. The two pyrene-based fluorophores were synthesized from the same starting material, so that the preparation of the beacons was simplified. The detection strategy of the beacons for target DNAs is based on "excimer-monomer emission switching" of the pyrene fluorophores: excimer emission of the pyrene moieties changed to monomer one when the beacons hybridized with the targets. This type of two-state mode of fluorescence allows unambiguous detection of the target DNAs because strict 1:1 correlation between the nonhybridized and the hybridized beacons can be monitored by the presence of isoemissive points of the fluorescence changes. The beacons can detect target 19-mer DNAs and can discriminate the targets from their single-nucleotide mismatches at 1 nM concentration. Advantages of the excimer-monomer switching molecular beacons were discussed in comparison with conventional ones.