Diastereomer Separation of Azobenzene-Tethered Oligodeoxyribonucleotides and Determination of Their Absolute Configurations by Enzymatic Digestion |
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| Authors: | Xingguo Liang Makoto Komiyama Hiroyuki Asanuma |
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| Institution: | 1. Department of Molecular Design and Engineering, Graduate School of Engineering , Nagoya University , Nagoya, Japan;2. Research Center for Advanced Science and Technology , The University of Tokyo , Tokyo, Japan;3. Department of Molecular Design and Engineering, Graduate School of Engineering , Nagoya University , Nagoya, Japan;4. CREST , Japan Science and Technology Agency (JST) , Kawaguchi, Japan |
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| Abstract: | Two diastereomers were produced by the introduction of azobenzene-tethering prochiral linker (2,2-bis(hydroxymethyl)propionic acid) in the modified ODN, which had been used for the photoregulation of DNA functions. We found that this modified ODN with sequence 5′-…pNpXpN…-3′ (p = phosphate; N = nucleoside; X = azobenzene residue) could be digested to pX (the phosphate at the 5′ side of X was left) by an over excess of Phosphodiesterase I. By comparing the retention time of pX from the separated diastereomer with that of authentic R- or S-pX on chiral HPLC, absolute configuration could be easily determined. |
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| Keywords: | Diastereomer separation modified oligodeoxynucleotide absolute configuration enzymatic digestion Phosphodiesterase |
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