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"Development Of A 'clickable' Non-natural Nucleotide To Visualize The Replication Of Non-instructional DNA Lesions." and the language of the book is English.


“Development Of A 'clickable' Non-natural Nucleotide To Visualize The Replication Of Non-instructional DNA Lesions.” Metadata:

  • Title: ➤  Development Of A 'clickable' Non-natural Nucleotide To Visualize The Replication Of Non-instructional DNA Lesions.
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  • Language: English

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  • Internet Archive ID: pubmed-PMC3300027

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This article is from <a href="//archive.org/search.php?query=journaltitle%3A%28Nucleic%20Acids%20Research%29" rel="nofollow">Nucleic Acids Research</a>, <a href="//archive.org/search.php?query=journaltitle%3A%28Nucleic%20Acids%20Research%29%20AND%20volume%3A%2840%29" rel="nofollow">volume 40</a>.<h2>Abstract</h2>The misreplication of damaged DNA is an important biological process that produces numerous adverse effects on human health. This report describes the synthesis and characterization of a non-natural nucleotide, designated 3-ethynyl-5-nitroindolyl-2′-deoxyriboside triphosphate (3-Eth-5-NITP), as a novel chemical reagent that can probe and quantify the misreplication of damaged DNA. We demonstrate that this non-natural nucleotide is efficiently inserted opposite an abasic site, a commonly formed and potentially mutagenic non-instructional DNA lesion. The strategic placement of the ethynyl moiety allows the incorporated nucleoside triphosphate to be selectively tagged with an azide-containing fluorophore using ‘click’ chemistry. This reaction provides a facile way to quantify the extent of nucleotide incorporation opposite non-instructional DNA lesions. In addition, the incorporation of 3-Eth-5-NITP is highly selective for an abasic site, and occurs even in the presence of a 50-fold molar excess of natural nucleotides. The biological applications of using 3-Eth-5-NITP as a chemical probe to monitor and quantify the misreplication of non-instructional DNA lesions are discussed.

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