Vol 51(2017) N 3 p. 474-482; DOI 10.1134/S0026893317030025
A.V. Chudinov1*, Y.Y. Kiseleva2, V.E. Kuznetsov1, V.E. Shershov1, M.A. Spitsyn1, T.O. Guseinov1, S.A. Lapa1, E.N. Timofeev1, A.I. Archakov2, A.V. Lisitsa2, S.P. Radko2, A.S. Zasedatelev1
Enzymatic synthesis of high-modified DNA1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia
2Orekhovich Institute of Biomedical Chemistry, Moscow, 119121 Russia
Received - 2016-05-10; Accepted - 2016-07-11
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.
modified nucleotides, deoxyuridine, primer extension, SELEX