Chudinov A V, Kiseleva Y Y, Kuznetsov V E, Shershov V E, Spitsyn M A, Guseinov T O, Lapa S A, Timofeev E N, Archakov A I, Lisitsa A V, Radko S P, Zasedatelev A S
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia.
Orekhovich Institute of Biomedical Chemistry, Moscow, 119121 Russia.
Mol Biol (Mosk). 2017 May-Jun;51(3):534-544. doi: 10.7868/S0026898417030028.
Here, we describe the synthesis and purification of six deoxyuridine triphosphate derivatives that contain protein-like functional groups and alkene linkers of various lengths. Using KOD XL and Deep Vent polymerases, these derivatives have been incorporated into single-stranded DNA, achieving a high degree of DNA modification. These polymerases are able to utilize highly modified DNA strands as templates for synthesizing unmodified DNA. The synthesized deoxyuridine triphosphate derivatives are promising as substrates for producing modified aptamers to various target proteins using, e.g., the systematic evolution of ligands by exponential enrichment (SELEX) methodology.
在此，我们描述了六种含有类蛋白质官能团和不同长度烯烃连接基的脱氧尿苷三磷酸衍生物的合成与纯化。使用KOD XL和Deep Vent聚合酶，这些衍生物已被掺入单链DNA中，实现了高度的DNA修饰。这些聚合酶能够利用高度修饰的DNA链作为模板来合成未修饰的DNA。所合成的脱氧尿苷三磷酸衍生物有望作为底物，用于通过例如指数富集配体系统进化（SELEX）方法来生产针对各种靶蛋白的修饰适配体。
