A MutS Protein-immobilized Au Electrode for Detecting Single-base Mismatch of DNA

HAN Aishan, TAKARADA Tohru, SHIBATA Taiki, NAKAYAMA Masamichi, MAEDA Mizuo

doi:10.2116/analsci.22.663

A MutS Protein-immobilized Au Electrode for Detecting Single-base Mismatch of DNA

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HAN Aishan

Department of Applied Chemistry, Graduate School of Engineering, Kyushu University
TAKARADA Tohru

Bioengineering Laboratory, RIKEN
SHIBATA Taiki

Department of Applied Chemistry, Graduate School of Engineering, Kyushu University
NAKAYAMA Masamichi

Department of Applied Chemistry, Graduate School of Engineering, Kyushu University
MAEDA Mizuo

Department of Applied Chemistry, Graduate School of Engineering, Kyushu University Bioengineering Laboratory, RIKEN

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タイトル別名

MutS Protein immobilized Au Electrode for Detecting Single base Mismatch of DNA

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A novel electrochemical biosensor was developed to detect gene mutation by using a DNA-mismatch binding protein: MutS from Escherichia coli. The MutS protein was immobilized onto an Au-electrode surface via complex formation between a histidine tag of the MutS protein and a thiol-modified nitrilotriacetic acid chemically adsorbed on the Au-electrode surface. When a target double-stranded DNA having a single-base mismatch was captured by the MutS protein on the electrode, some electrostatic repulsion arose between polyanionic DNA strands and anionic redox couple ions. Consequently, their redox peak currents on a cyclic voltammogram with the Au electrode drastically decreased, depending on the concentration of the target DNA, according to the redox couple-mediated artificial ion-channel principle. By using this assay, one can detect all types of single-base mismatch and single-base deletion.