PMID:12903217

Citation	*Han, A, Shibata, T, Takarada, T and Maeda, M' (2002) Gene mutation assay using a MutS protein-modified electrode.* Nucleic Acids Res. Suppl. '**:287-8
Abstract	A novel electrochemical biosensor for gene mutation detection was developed using a DNA mismatch recognizing protein MutS from E. coli. The MutS protein was immobilized onto an Au electrode by coordination of His-tag at its C-terminus to vacant sites of Ni(II)-nitrilotriacetato complex attached to the surface of electrode. When a target DNA duplex having a mismatch site was captured by the MutS protein on the electrode, the electrostatic repulsion arose between polyanionic DNA duplexes and negatively-charged ferrocyanide/ferricyanide 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. Using this assay, we could detect GT mismatch and deletion mutation in the double-stranded DNA.
Links	PubMed
Keywords	Adenosine Triphosphatases/chemistry; Bacterial Proteins; DNA-Binding Proteins; Electrodes; Escherichia coli Proteins/chemistry; MutS DNA Mismatch-Binding Protein; Mutation
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