Fluorescent sensor for mercury
View Patent ↗The present invention provides a sensor for detecting mercury, comprising: a first polynucleotide, comprising a first region, and a second region, a second polynucleotide, a third polynucleotide, a fluorophore, and a quencher, wherein the third polynucleotide is optionally linked to the second region; the fluorophore is linked to the first polynucleotide and the quencher is linked to the second polynucleotide, or the fluorophore is linked to the second polynucleotide and the quencher is linked to the first polynucleotide; the first region and the second region hybridize to the second polynucleotide; and the second region binds to the third polynucleotide in the presence of Hg 2+ ions.
1. A sensor for detecting mercury, comprising:
a first polynucleotide, comprising a first region, and a second region,
a second polynucleotide,
a third polynucleotide,
a fluorophore, and
a quencher,
wherein the third polynucleotide is optionally linked to the second region;
the fluorophore is linked to the first polynucleotide and the quencher is linked to the second polynucleotide, or the fluorophore is linked to the second polynucleotide and the quencher is linked to the first polynucleotide;
when the first region and the second region hybridize to the second polynucleotide, the quencher quenches the fluorophore; and
the second region binds to the third polynucleotide when in the presence of Hg 2+ ions, releasing the second polynucleotide so that the quencher does not quench the fluorophore.
2. The sensor of claim 1 , wherein the fluorophore is linked to the 5′ end of the first polynucleotide.
3. The sensor of claim 1 , wherein the quencher is linked to the 3′ end of the second polynucleotide.
4. The sensor of claim 1 , wherein the fluorophore is linked to the 3′ end of the second polynucleotide.
5. The sensor of claim 1 , wherein the quencher is linked to the 5′ end of the first polynucleotide.
6. The sensor of claim 1 , wherein the fluorophore is FAM.
7. The sensor of claim 1 , wherein the quencher is non-flourescent quenchers.
8. The sensor of claim 1 , wherein the first polynucleotide comprises the nucleic-acid sequence of SEQ ID NO:1.
9. The sensor of claim 1 , wherein the second polynucleotide comprises the nucleic acid sequence of SEQ ID NO:2.
10. The sensor of claim 1 , wherein the the second region and the third polynucleotide form 2 to 20 thymine-thymine mismatches.
11. The sensor of claim 1 , wherein the second region and the third polynucleotide form 4 to 10 thymine-thymine mismatches.
12. The sensor of claim 1 , wherein the second region and the third polynucleotide form 5 to 8 thymine-thymine mismatches.
13. A method of detecting the presence of mercury in a sample, comprising:
(a) forming a mixture comprising the sample and a sensor comprising:
a first polynucleotide, comprising a first region, and a second region,
a second polynucleotide,
a third polynucleotide,
a fluorophore, and
a quencher,
wherein the third polynucleotide is optionally linked to the second region;
the fluorophore is linked to the first polynucleotide and the quencher is linked to the second polynucleotide, or the fluorophore is linked to the second polynucleotide and the quencher is linked to the first polynucleotide;
when the first region and the second region hybridize to the second polynucleotide, the quencher quenches the fluorophore; and
the second region binds to the third polynucleotide when in the presence of Hg 2+ ions, releasing the second polynucleotide so that the quencher does not quench the fluorophore, and
(b) measuring the fluorescence of the mixture.
14. The method of claim 13 , wherein the fluorophore is linked to the 5′ end of the first polynucleotide.
15. The method of claim 13 , wherein the quencher is linked to the 3′ end of the second polynucleotide.
16. The method of claim 13 , wherein the fluorophore is linked to the 3′ end of the second polynucleotide.
17. The method of claim 13 , wherein the quencher is linked to the 5′ end of the first polynucleotide.
18. The method of claim 13 , wherein the fluorophore is FAM.
19. The method of claim 13 , wherein the quencher is non-flourescent quenchers.
20. The method of claim 13 , wherein the second region and the third polynucleotide form 4 to 10 thymine-thymine mismatches.
21. The method of claim 13 , wherein the sample comprises a water sample.
22. The method of claim 13 , wherein the sample comprises a bodily fluid.
23. The method of claim 13 , wherein an increase in fluorescence is indicative of the presence of mercury.
24. A method of determining the concentration of mercury in a sample, comprising:
(a) forming a mixture comprising the sample and a sensor comprising:
a first polynucleotide, comprising a first region, and a second region,
a second polynucleotide,
a third polynucleotide,
a fluorophore, and
a quencher,
wherein the third polynucleotide is optionally linked to the second region;
the fluorophore is linked to the first polynucleotide and the quencher is linked to the second polynucleotide, or the fluorophore is linked to the second polynucleotide and the quencher is linked to the first polynucleotide;
when the first region and the second region hybridize to the second polynucleotide, the quencher quenches the fluorophore; and
the second region binds to the third polynucleotide when in the presence of Hg 2+ ions, releasing the second polynucleotide so that the quencher does not quench the fluorophore,
(b) measuring the fluorescence of the mixture; and
(c) comparing the measurement obtained in (b) with that of a calibration curve created using known concentrations of mercury.
25. The method of claim 24 , wherein the second region and the third polynucleotide form 4 to 10 thymine-thymine mismatches.
26. The method of claim 24 , wherein the sample comprises a water sample.
27. The method of claim 24 , wherein the sample comprises a bodily fluid.