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多自旋标记DNA的酶促合成

Enzymatic synthesis of multi spin-labeled DNA.

作者信息

Obeid Samra, Yulikov Maxim, Jeschke Gunnar, Marx Andreas

机构信息

Department of Chemistry and Konstanz Research School Chemical Biology, University of Konstanz, 78457, Konstanz, Germany.

出版信息

Nucleic Acids Symp Ser (Oxf). 2008(52):373-4. doi: 10.1093/nass/nrn188.

Abstract

Electron paramagnetic resonance (EPR) spectroscopy was applied extensively in studies of nucleic acid structures and dynamics. Two modified 2'-deoxyuridine triphosphates were synthesized bearing a spin label linked to the base by a rigid linker to ensure a tight coupling of spin label dynamics. The incorporation of both spin-labeled nucleotides could be shown in primer extension reactions in presence of DNA polymerases from eukaryotic, prokaryotic, and archaic origin. In further experiments we were able to introduce multiple spin labels during primer extension reactions.

摘要

电子顺磁共振(EPR)光谱在核酸结构与动力学研究中得到了广泛应用。合成了两种经修饰的三磷酸2'-脱氧尿苷,它们带有通过刚性连接基团与碱基相连的自旋标记,以确保自旋标记动力学的紧密耦合。在真核、原核和古生菌来源的DNA聚合酶存在的情况下,引物延伸反应中均可显示两种自旋标记核苷酸的掺入。在进一步的实验中,我们能够在引物延伸反应过程中引入多个自旋标记。

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