通过点击化学将血红素共价连接到形成 G-四链体的寡核苷酸上来增强过氧化物酶模拟 DNA 酶的活性。

Enhancement of Activity for Peroxidase-Mimicking DNAzyme by Covalent Attachment of Hemin to G-Quadruplex-Forming Oligonucleotide Using Click Chemistry.

机构信息

Department of Bioanalytical Chemistry, Faculty of Chemistry, Adam Mickiewicz University, Poznań, Poland.

出版信息

Methods Mol Biol. 2022;2439:291-299. doi: 10.1007/978-1-0716-2047-2_18.

DOI:10.1007/978-1-0716-2047-2_18

Abstract

Peroxidase-mimicking DNAzymes are used in the development of new bioanalytical assays due to their advantages like thermal stability, simple synthesis and purification, and ability to hybridize with the complementary strand of nucleic acid. Here, we describe the method of covalent attachment of hemin to DNA oligonucleotide using click chemistry that allows good yield (60-70%) of the final conjugate product. The activity of obtained DNAzymes is monitored using chromogenic and fluorogenic substrates.

摘要

过氧化物酶模拟 DNA 酶由于其具有热稳定性、合成和纯化简单以及能够与核酸互补链杂交等优点，被用于新型生物分析测定法的开发。在此，我们描述了使用点击化学将血红素共价连接到 DNA 寡核苷酸上的方法，该方法可使最终缀合物产物的收率（60-70%）良好。通过显色和荧光底物来监测获得的 DNA 酶的活性。