Gene mutation assay using a MutS protein-modified electrode.
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.
Han
Aishan
A
Graduate School of Engineering, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
Shibata
Taiki
T
Takarada
Tohru
T
Maeda
Mizuo
M
D000251:Adenosine Triphosphatases / Q000737:chemistry*; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D004566:Electrodes; D029968:Escherichia coli Proteins / Q000737:chemistry*; D051719:MutS DNA Mismatch-Binding Protein; D009154:Mutation
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D000251:Adenosine Triphosphatases; D051719:MutS DNA Mismatch-Binding Protein; C497175:MutS protein, E coli