Probing DNA sequences in solution with a monomer-excimer fluorescence color change
Probing DNA sequences in solution with a monomer-excimer fluorescence color change
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DOI：
10.1093/nar/26.16.3789
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发表时间：
1998-08-15
影响因子：
14.9
通讯作者：
Kool, ET
中科院分区：
文献类型：
--
作者：
Paris, PL;Langenhan, JM;Kool, ET
本文描述了一种简单的荧光核苷类似物在通过溶液杂交检测点突变中的应用。通过脱氧核糖的亚磷酰胺衍生物将芘置于一对寡脱氧核苷酸探针的3'和5'末端，该荧光团连接在1'位置，取代DNA碱基。含有该荧光团的互补靶序列的双探针的相邻结合导致从蓝色芘单体发射（λ（max）= 381 - 398 nm）到绿白色受激准分子发射（λ（max）= 490 nm）的显著光谱变化。两个探针的相对结合位置的优化表明，当它们与部分末端重叠结合时，发生最大的光谱变化。在最佳取向，探针的单体发射带的强度降低了多达7倍，准分子带增加了40倍，结合互补的目标。应用到检测的单碱基点突变的解决方案。
The use of a simple fluorescent nucleoside analogue in detection of point mutations by hybridization in solution is described. Pyrene is placed at 3' and 5' ends of a pair of oligodeoxynucleotide probes via a phosphoramidite derivative of deoxyribose with this fluorophore attached at the 1' position, replacing a DNA base. Adjacent binding of dual probes containing this fluorophore to a complementary target sequence results in a pronounced spectral change from blue pyrene monomer emission (lambda(max) = 381 398 nm) to green-white excimer emission (lambda(max) = 490 nm). Optimization of the relative binding positions of the two probes shows that the greatest spectral change occurs when they bind with partial end overlap. In optimum orientation, the monomer emission band for the probes decreases intensity by as much as a factor of seven and the excimer band increases up to 40-fold on binding a complementary target. Application to the detection of a single-base point mutation in solution is described.