- METHOD FOR DETECTING MISMATCHED DNA USING FLUORESCENCE CHANGE OF MutS MUTANT-FLUOROPHORE CONJUGATE

摘要

A method for detecting a mismatched DNA by using fluorescence change of the MutS mutant-fluorophore conjugate is provided to improve simplicity and convenience of detection compared to the conventional methods including gel mobility shift assay and a method employing MutS immobilized on the surface. The mismatched DNA is detected by adding a sample DNA into the MutS mutant-fluorophore conjugate and measuring the fluorescence change of the conjugate, wherein the MutS mutant-fluorophore conjugate is prepared by substituting a 42th amino acid cysteine by alanine and a 469th amino acid threonine by cysteine in the MutS protein of Thermus aquaticus, Taq., and binding a thio-specific fluorophore to the mutated 469th amino acid residue; and the thio-specific fluorophore is IAANS(2-(4'-(iodoacetoamido)aniline)naphthalene-6-sulfonic acid), BADAN(6-bromoacetyl-2-dimethylamino naphthalene) or N-(1-pyrene) iodoacetamide.