Method for detecting target nucleotide sequences
A method for detecting nucleic acids wherein the binding activity of a nucleic acid aptamer, which is formed by the hybridization of a target nucleotide sequence and a probe, is detected using the binding with a ligand as an index has been provided. Utilizing a structure such as the three-way junction, whose binding activity significantly changes depending on a mismatch of a single nucleotide, as the nucleic acid aptamer SNP can be detected.
1. A method for detecting a target nucleotide sequence, which comprises the steps of:
(a) hybridizing a probe to the target nucleotide sequence to form a nucleic acid aptamer having a ligand-binding affinity, wherein the ligand is cholic acid, and wherein the aptamer has three stems and binds with the ligand at the position where the three stems intersect; and
(b) detecting the presence of the target nucleotide sequence using the ligand-binding affinity of the aptamer as an index.
2. The method of claim 1 , wherein the target nucleotide sequence contains a single nucleotide polymorphism, and the affinity of the formed aptamer for the cholic acid ligand varies depending on the substitution of the single nucleotide in the target molecule.
3. The method of claim 1 , wherein at least two of three base pairs at the position where the three stems intersect are G-C nucleotide pairs.
4. The method of claim 3 , wherein each of the three stems is longer than three base pairs.