Diene-modified nucleotides for the DielsAlder-mediated functional tagging of DNA

被引：62

作者：

Borsenberger, Vinciane ^{[1
]}

Howorka, Stefan ^{[1
]}

机构：

[1] UCL, Inst Struct Mol Biol, Dept Chem, London WC1H 0AJ, England

来源：

NUCLEIC ACIDS RESEARCH | 2009年 / 37卷 / 05期

基金：

英国工程与自然科学研究理事会;

关键词：

ALDER; OLIGONUCLEOTIDES; 5'-TRIPHOSPHATES; CYCLOADDITION; NUCLEOSIDES; PROTEIN; ACIDS;

D O I：

10.1093/nar/gkn1066

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We explore the potential of the DielsAlder cycloaddition for the functional tagging of DNA strands. A deoxyuridine triphosphate derivative carrying a diene at position 5 of the pyrimidine base was synthesized using a two-step procedure. The derivative was efficiently accepted as substrate in enzymatic polymerization assays. Diene carrying strands underwent successful cycloaddition with maleimide-terminated fluorescence dyes and a polymeric reagent. Furthermore, a nucleotide carrying a peptide via a DielsAlder cyclohexene linkage was prepared and sequence-specifically incorporated into DNA. The DielsAlder reaction presents a number of positive attributes such as good chemoselectivity, water compatibility, high-yield under mild conditions and no additional reagents apart from a diene and a dienophile. Furthermore, suitable dienophiles are commercially available in the form of maleimide-derivatives of fluorescent dyes and bioaffinity tags. Based on these advantages, diene- and cyclohexene-based nucleotide triphosphates are expected to find wider use in the area of nucleic acid chemistry.

引用

页码：1477 / 1485

页数：9

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