Use of a novel 5'-regioselective phosphitylating reagent for one-pot synthesis of nucleoside 5'-triphosphates from unprotected nucleosides.
Use of a novel 5'-regioselective phosphitylating reagent for one-pot synthesis of nucleoside 5'-triphosphates from unprotected nucleosides.
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DOI：
10.1002/0471142700.nc0130s52
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发表时间：
2013-03
影响因子：
--
通讯作者：
Huang, Zhen
中科院分区：
文献类型：
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作者：
Caton-Williams, Julianne;Hoxhaj, Rudiona;Fiaz, Bilal;Huang, Zhen
5′-三磷酸是DNA和RNA酶促合成的基础。本单元介绍了一种方便合成2′-脱氧核糖和核糖核苷5′-三磷酸（dNTPs和NTPs）的方法，其中包含所有天然碱基和修饰碱基。这种一锅法合成也可用于制备三磷酸酯类似物，所述三磷酸酯类似物含有取代三磷酸酯的α磷酸酯的非桥氧原子的硫或硒原子。这些S-或Se-修饰的dNTP和NTP可用于制备非对映体纯的硫代磷酸酯核酸（PS-NA）或硒代磷酸酯核酸（PSe-NA，即，一种硒衍生的核酸：SeNA）。即使没有大量纯化，合成的dNTP或NTP也具有高质量，并且可以直接用于DNA聚合或RNA转录。本方案单元描述了5′-三磷酸的合成和纯化、天然核酸和硫或硒修饰核酸的分析和确认。
The 5′-triphosphates are the building blocks for the enzymatic synthesis of DNAs and RNAs. This unit presents a protocol for the convenient synthesis of 2′-deoxyribo- and ribo-nucleoside 5′-triphosphates (dNTPs and NTPs) containing all the natural bases and the modified bases. This one-pot synthesis can also be applied to prepare the triphosphate analogs that contain sulfur or selenium atoms replacing the non-bridging oxygen atoms of the alpha phosphates of the triphosphates. These S- or Se-modified dNTPs and NTPs can be used to prepare diastereomerically-pure phosphorothioate nucleic acids (PS-NAs) or phosphoroselenoate nucleic acids (PSe-NAs, i.e., one type of selenium-derivatized nucleic acids: SeNA). Even without extensive purification, the synthesized dNTPs or NTPs are of high quality and can be directly used in DNA polymerization or RNA transcription. Synthesis and purification of the 5′-triphosphates, analysis and confirmation of natural and sulfur-or selenium-modified nucleic acids are described in this protocol unit.