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通过含有 5-乙炔基尿嘧啶的 DNA 的铜AAC 点击反应来关闭细菌 RNA 聚合酶的转录。

Turning Off Transcription with Bacterial RNA Polymerase through CuAAC Click Reactions of DNA Containing 5-Ethynyluracil.

机构信息

Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Flemingovo nam. 2, 16610, Prague 6, Czech Republic.

Institute of Microbiology, Czech Academy of Sciences, 14220, Prague 4, Czech Republic.

出版信息

Chemistry. 2018 Jun 12;24(33):8311-8314. doi: 10.1002/chem.201801757. Epub 2018 May 17.

Abstract

Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) click reaction in the major groove of DNA containing 5-ethynyluracil (U ) with azides was used for turning off sequence-specific protein-DNA interactions. The concept was first demonstrated on switching off cleavage of short modified DNA by restriction endonuclease BamHI-HF. Finally, DNA template containing U was used for in vitro transcription with E. coli RNA polymerase and the transcription was turned off by CuAAC with 3-azidopropane-1,2-diol or 3-azido-7-hydroxycoumarin.

摘要

铜(I)催化的叠氮化物-炔烃环加成(CuAAC)点击反应在含有 5-乙炔基尿嘧啶(U)的 DNA 大沟中与叠氮化物反应,用于关闭序列特异性蛋白-DNA 相互作用。该概念首先通过限制内切酶 BamHI-HF 证明了在关闭短修饰 DNA 的切割方面的应用。最后,使用含有 U 的 DNA 模板进行大肠杆菌 RNA 聚合酶的体外转录,并用 3-叠氮丙烷-1,2-二醇或 3-叠氮-7-羟基香豆素通过 CuAAC 关闭转录。

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