ACCELERATED HYBRIDIZATION OF OLIGONUCLEOTIDES TO DUPLEX DNA
ACCELERATED HYBRIDIZATION OF OLIGONUCLEOTIDES TO DUPLEX DNA
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DOI：
10.1074/jbc.270.24.14712
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发表时间：
1995-06-16
影响因子：
4.8
通讯作者：
COREY, DR
中科院分区：
文献类型：
--
作者：
IYER, M;NORTON, JC;COREY, DR
我们报告了两种加速寡核苷酸与DNA杂交的策略。我们证明了低聚脱氧核糖核苷酸和肽核酸低聚物通过d环形成杂交到双工DNA内的反向重复序列。寡核苷酸与双模板形成活性复合物，T7 DNA聚合酶可识别其进行初始聚合。聚合产物的定量可以估计杂交速率，并且肽核酸低聚物和寡核苷酸-蛋白加合物的结合常数分别比类似的未修饰寡核苷酸高500倍和12000倍。总之，这些结果表明，双工DNA中的序列可以被沃森-克里克碱基配对靶向，化学修饰可以显著提高链结合的速度。这些发现应该有助于靶向低聚物引发DNA聚合，检测诊断序列，以及基因表达的中断。观察到的杂交加速可能为研究RecA或其他蛋白质加速链入侵的能力提供了新的视角。
We report two strategies for accelerating the hybridization of oligonucleotides to DNA. We demonstrate that oligodeoxyribonucleotides and peptide nucleic acid oligomers hybridize to inverted repeats within duplex DNA by D-loop formation. Oligonucleotides and duplex template form an active complex, which can be recognized by T7 DNA polymerase to prime polymerization. Quantitation of polymerization products allowed the rate of hybridization to be estimated, and peptide nucleic acid oligomers and oligonucleotide-protein adducts anneal with association constants 500- and 12,000-fold greater, respectively, than the analogous unmodified oligonucleotides. Together, these results indicate that sequences within duplex DNA can be targeted by Watson-Crick base pairing and that chemical modifications can dramatically enhance the rate of strand association. These findings should facilitate targeting of oligomers for priming DNA polymerization, the detection of diagnostic sequences, and the disruption of gene expression. The observed acceleration of hybridization may offer a new perspective on the ability of RecA or other proteins to accelerate strand invasion.