NEW TOOLS FOR POINT MUTATIONS DETECTION
In the present research we designed novel fluorescent probes based on 5'-monopyrene derivatives of oligo(2'-O-methylribonucleotides) and tandems of 5'- and 3'-monopyrene derivatives of oligo(2'-O-methylribonucleotides) targeting to the fragment of methylenetetrahydrofolatereductase (MTHFR) gene containing C677T polymorphism. The effect of probes length, structure of linkers connecting pyrene residue with oligonucleotide part and the presence of thymidine attached at the 3'-termini by a 3'-3'-phosphodiester internucleotide bond on fluorescent properties of probes and their duplexes with DNA targets was studied. The fluorescent properties of proposed probes were shown to be sensitive to the mononucleotide substitution in DNA target. The developed probes were demonstrated to be utilized for the detection of polymorphism C677T in MTHFR gene by the method of thermal duplex denaturation with fluorescence change registration.