Publication details

Sensitive electrochemical determination of unlabeled MutS protein and detection of point mutations in DNA

Authors

PALEČEK Emil MASAŘÍK Michal KIZEK René KUHLMEIER Dirk HASSMANN Jörg SCHÜLEIN Jürgen

Year of publication 2004
Type Article in Periodical
Magazine / Source Analytical Chemistry
MU Faculty or unit

Faculty of Science

Citation
Field Genetics and molecular biology
Keywords MEDIATED CHARGE-TRANSPORT; MODIFIED ELECTRODES; NUCLEIC-ACIDS; STRIPPING VOLTAMMETRY; BIOACTIVE PEPTIDES; MERCURY-ELECTRODES; CARBON ELECTRODES; MISMATCH REPAIR; SOLID AMALGAMS; HYBRIDIZATION
Description MutS protein plays an important role in the DNA repair system in prokaryotic and eukaryotic cells; it recognizes unpaired and mispaired bases in duplex DNA and can be used for detection of point mutations in vitro. We have shown that small amounts of this protein can be detected electrochemically at mercury and carbon electrodes without any labeling. Using constant current stripping analysis (CPSA) and mercury electrodes, tens of attomoles of this protein can be detected. The sensitivity of the determination at carbon electrodes is by more than 3 orders of magnitude lower. Using biotinylated DNA duplexes attached to magnetic beads, single-base mismatches and insertion/deletions were recognized by MutS. Picogram amounts of this protein were detected by CPSA after MutS releasing from the beads.

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