Röthlisberger Pascal, Levi-Acobas Fabienne, Hollenstein Marcel
Laboratory for Bioorganic Chemistry of Nucleic Acids, Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris Cedex 15, France; CNRS UMR3523 Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris Cedex 15, France.
Laboratory for Bioorganic Chemistry of Nucleic Acids, Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris Cedex 15, France; CNRS UMR3523 Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris Cedex 15, France.
Bioorg Med Chem Lett. 2017 Feb 15;27(4):897-900. doi: 10.1016/j.bmcl.2017.01.009. Epub 2017 Jan 6.
5-Ethynyl-2'-deoxyuridine is a common base-modified nucleoside analogue that has served in various applications including selection experiments for potent aptamers and in biosensing. The synthesis of the corresponding triphosphates involves a mild acidic deprotection step. Herein, we show that this deprotection leads to the formation of other nucleoside analogs which are easily converted to triphosphates. The modified nucleoside triphosphates are excellent substrates for numerous DNA polymerases under both primer extension and PCR conditions and could thus poison selection experiments by blocking sites that need to be further modified. The formation of these nucleoside analogs can be circumvented by application of a new synthetic route that is described herein.
5-乙炔基-2'-脱氧尿苷是一种常见的碱基修饰核苷类似物，已应用于各种领域，包括高效适配体的筛选实验和生物传感。相应三磷酸酯的合成涉及温和的酸性脱保护步骤。在此，我们表明这种脱保护会导致形成其他核苷类似物，这些类似物很容易转化为三磷酸酯。在引物延伸和聚合酶链式反应（PCR）条件下，修饰的核苷三磷酸酯都是众多DNA聚合酶的优良底物，因此可能会通过阻断需要进一步修饰的位点而干扰筛选实验。本文描述的一种新合成路线可以避免这些核苷类似物的形成。
