In Vitro Analysis of DNA-Protein Interactions in Gene Transcription Using DNAzyme-Based Electrochemical Assay

被引：24

作者：

Li, Chao ^{[1
,2
]}

Tao, Yaqin ^{[1
,2
]}

Yang, Yi ^{[1
,2
]}

Xiang, Yang ^{[1
,2
]}

Li, Genxi ^{[1
,2
,3
]}

机构：

[1] Nanjing Univ, Dept Biochem, State Key Lab Pharmaceut Biotechnol, Nanjing 210093, Jiangsu, Peoples R China

[2] Nanjing Univ, Dept Biochem, Collaborat Innovat Ctr Chem Life Sci, Nanjing 210093, Jiangsu, Peoples R China

[3] Shanghai Univ, Sch Life Sci, Ctr Mol Recognit & Biosensing, Shanghai 200444, Peoples R China

来源：

ANALYTICAL CHEMISTRY | 2017年 / 89卷 / 09期

基金：

中国国家自然科学基金;

关键词：

BINDING; POLYMERASE; BEACONS;

D O I：

10.1021/acs.analchem.7b00329

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

The interaction between protein and DNA elements controls a variety of functions of genomes. The development of a convenient and cost-effective method for investigating the sequence specificity of DNA-binding proteins represents an important challenge. In response, we have introduced an electrochemical assay in this work for specific and sensitive analysis of interaction between protein and nucleic acid in nucleic extracts, based on the protein-induced distinctive motion behavior of DNA deoxyribozyme (DNAzyme) on an electrode surface. As a proof of principle, we have also presented assays for the rapid, sensitive, and selective detection of three transcription factors (NF-kappa B, SP6 RNA polymerase, and HNF-4 alpha), as well as the analysis of binding affinity of the mutated protein-binding sequence, and even screening of the binding sequence of HNF-4 alpha protein in vitro. This work may open new opportunity for in-depth profiling of the sequence specificity of DNA-binding proteins and study of nucleotide polymorphisms in known protein-binding sites.

引用

页码：5003 / 5007

页数：5