Electrochemical detection of mismatched DNA using a MutS probe.
A direct and label-free electrochemical biosensor for the detection of the protein-mismatched DNA interaction was designed using immobilized N-terminal histidine tagged Escherichia coli. MutS on a Ni-NTA coated Au electrode. General electrochemical methods, cyclic voltammetry (CV), electrochemical quartz crystal microbalance (EQCM) and impedance spectroscopy, were used to ascertain the binding affinity of mismatched DNAs to the MutS probe. The direct results of CV and impedance clearly reveal that the interaction of MutS with the CC heteroduplex was much stronger than that with AT homoduplex, which was not differentiated in previous results (GT > CT > CC approximately AT) of a gel mobility shift assay. The EQCM technique was also able to quantitatively analyze MutS affinity to heteroduplexes.
Cho
Minseon
M
Department of Chemistry, Pohang University of Science and Technology, Pohang, Gyungbuk, 790-784, South Korea.
Lee
Sohyun
S
Han
Se-Young
SY
Park
Jin-Young
JY
Rahman
Md Aminur
MA
Shim
Yoon-Bo
YB
Ban
Changill
C
D020137:Base Pair Mismatch; D015374:Biosensing Techniques / Q000295:instrumentation; D004247:DNA / Q000737:chemistry; D004563:Electrochemistry; D004566:Electrodes; D029968:Escherichia coli Proteins / Q000737:chemistry; D020180:Heteroduplex Analysis / Q000379:methods*; D015335:Molecular Probes / Q000737:chemistry; D051719:MutS DNA Mismatch-Binding Protein / Q000737:chemistry; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry
D029968:Escherichia coli Proteins; D015335:Molecular Probes; D009692:Nucleic Acid Heteroduplexes; D004247:DNA; D051719:MutS DNA Mismatch-Binding Protein; C497175:MutS protein, E coli