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Induction of binding sites for RecA aptamers by differentiated-competition capillary Electrophoresis-SELEX

文献类型： 外文期刊

作者： Yang, Ge 1 ; Liu, Wenjing 3 ; Zhao, Yi 2 ; Jiang, Guangyu 1 ; Zhu, Chao 4 ; Qu, Feng 2 ;

作者机构： 1.Chinese Acad Med Sci & Peking Union Med Coll, CAMS Key Lab Antiviral Drug Res, Beijing Key Lab Antimicrobial Agents, NHC Key Lab Biotechnol Antibiot,Inst Med Biotechn, Beijing 100050, Peoples R China

2.Beijing Inst Technol, Sch Life Sci, Key Lab Mol Med & Biotherapy, 5 South Zhongguancun St, Beijing 100081, Peoples R China

3.Capital Med Univ, Beijing Chest Hosp, Beijing TB & Thorac Tumor Res Inst, Beijing Key Lab Drug Resistance TB Res, Beijing, Peoples R China

4.Shandong Acad Agr Sci, Inst Qual Stand & Testing Technol Agroprod, Jinan 250100, Peoples R China

5.Shandong Prov Key Lab Test Technol Food Qual & Saf, Jinan 250100, Peoples R China

关键词： Aptamer; RecA; Capillary electrophoresis; CE-SELEX; Competitive binding

期刊名称：TALANTA （ 影响因子：6.1； 五年影响因子：5.4 ）

ISSN： 0039-9140

年卷期： 2024 年 267 卷

页码：

收录情况： SCI

摘要： The binding site-oriented SELEX strategy is an effective way to promote the functional evolution of aptamers. Here we report a novel aptamer screening strategy (Differentiated-competition Capillary Electrophoresis-SELEX), which enables candidate aptamers to be directionally induced to bind to different active sites of RecA. In this strategy, we introduce two competing binding factors into the "binding - dissociation" dynamic equilibrium system of RecA and ssDNA libraries. Due to the completely different binding mechanism of the competitive factor and the ssDNA library, it exerts different interference on the binding of RecA and the ssDNA library, which directed the binding site of aptamer candidates during the SELEX process. Multifunctional aptamers with high affinity and specificity were found to inhibit RecA activity by binding to different active sites. In conclusion, the SELEX method developed in our current study have identified a variety of biologically functional aptamers with relatively well-defined binding sites that regulate RecA protein activity, which has potential applications and future prospects for accurate screening of biological functional aptamers.

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