Enzymatic Incorporation of Modified Purine Nucleotides in DNA |
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| Authors: | Rania Abu?El?Asrar Lia Margamuljana Dr Mikhail Abramov Dr Omprakash Bande Dr Stefano Agnello Dr Miyeon Jang Prof?Dr Piet Herdewijn |
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| Affiliation: | 1. Medicinal Chemistry, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium;2. Present address: Institute of Chemistry and Biotechnology, Center for Organic and Medicinal Chemistry, Zurich University of Applied Sciences (ZHAW), W?denswil, Switzerland |
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| Abstract: | A series of nucleotide analogues, with a hypoxanthine base moiety (8‐aminohypoxanthine, 1‐methyl‐8‐aminohypoxanthine, and 8‐oxohypoxanthine), together with 5‐methylisocytosine were tested as potential pairing partners of N8‐glycosylated nucleotides with an 8‐azaguanine or 8‐aza‐9‐deazaguanine base moiety by using DNA polymerases (incorporation studies). The best results were obtained with the 5‐methylisocytosine nucleotide followed by the 1‐methyl‐8‐aminohypoxanthine nucleotide. The experiments demonstrated that small differences in the structure (8‐azaguanine versus 8‐aza‐9‐deazaguanine) might lead to significant differences in recognition efficiency and selectivity, base pairing by Hoogsteen recognition at the polymerase level is possible, 8‐aza‐9‐deazaguanine represents a self‐complementary base pair, and a correlation exists between in vitro incorporation studies and in vivo recognition by natural bases in Escherichia coli, but this recognition is not absolute (exceptions were observed). |
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| Keywords: | base-pair recognition DNA incorporation nucleic acids nucleosides synthetic biology |
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