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DNA colorimetric logic gate construction method based on metal ion regulation and control of exonuclease III shearing action

A technology of exonuclease and metal ions, applied in the field of molecular computers, can solve the problems of insufficient sensitive and complete logic system, inapplicability of in-situ detection, etc., and achieve good selectivity

Inactive Publication Date: 2015-08-05
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned detection methods of metal ions and the construction methods of logic gates require multi-step operations and high-end instruments and equipment, which are not suitable for in-situ detection and are essential for building a sensitive and complete logic system. not enough

Method used

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  • DNA colorimetric logic gate construction method based on metal ion regulation and control of exonuclease III shearing action
  • DNA colorimetric logic gate construction method based on metal ion regulation and control of exonuclease III shearing action
  • DNA colorimetric logic gate construction method based on metal ion regulation and control of exonuclease III shearing action

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Construction of OR logic gate

[0027] The basic principles of DNA logic gate construction are as follows figure 1 Shown. Mix 20 μL of DNA 1 and 2 hybridization solution with 20 μL of DNA 3 and 4 hybridization solution to prepare quadruplicate solutions. Add 20 μL of water to the above solution to (0,0) state, add 10 μL of Ag + (5 μM) is (0,1) state, add 10 μL Hg 2+ (5 μM) is (1,0) state, and 10 μL Hg is added at the same time 2+ (5 μM) and 10 μL Ag + (5 μM) is the (1,1) state. Add 10 μL Exo Ш to each sample and incubate at 37 °C for 30 minutes. Then, add 20 μL of signal probe 1 to each sample to form a G-quadruplex, and then add 10 μL of a HEPES buffer solution containing 2 μM hemin, and react at room temperature for 1 h to form a G-quadruplex-hemorin Vegetarian complex. Finally, add 15 μL of TMB-H to each sample 2 O 2 Solution, G-quadruplex-heme catalyzes TMB color development. After 4 minutes of reaction, 200 μL of 2 M sulfuric acid is added to terminate the catalyti...

Embodiment 2

[0029] AND logic gate construction

[0030] Hybridize 20 μL of DNA 5 and 6 to prepare quadruplicate solutions. Add 20 μL of water to the above solution for (0,0) state; add 10 μL of Ag + (5 μM) is (0,1) state, add 10 μL Hg 2+ (5 μM) is (1,0) state, and 10 μL Hg is added at the same time 2+ (5 μM) and 10 μL Ag + (5 μM) is the (1,1) state. Add 10 μL Exo Ш to each sample and incubate at 37 °C for 30 minutes. Then, add 20 μL of signal probe 1 to each sample to form a G-quadruplex, and then add 10 μL of a HEPES buffer solution containing 2 μM hemin, and react at room temperature for 1 h to form a G-quadruplex-hemorin Vegetarian complex. Finally, add 15 μL of TMB-H to each sample 2 O 2 Solution, G-quadruplex-heme catalyzes TMB color development. After 4 minutes of reaction, 200 μL of 2 M sulfuric acid is added to terminate the catalytic reaction. Characterize the constructed logic gate using ultraviolet-visible spectroscopy, and the results are as follows image 3 Shown. The absorb...

Embodiment 3

[0032] Construction of INHIBIT logic gate

[0033] Mix 20 μL of DNA 1 and 2 hybridization solution with 20 μL of DNA 7 and 8 hybridization solution to prepare quadruplicate solutions. Add 20 μL of water to the above solution to (0,0) state, add 10 μL of Ag + (5 μM) is (0,1) state, add 10 μL Hg 2+ (5 μM) is (1,0) state, and 10 μL Hg is added at the same time 2+ (5 μM) and 10 μL Ag + (5 μM) is the (1,1) state. Add 10 μL Exo Ш to each sample and incubate at 37 °C for 30 minutes. Then, add 20 μL of signal probe 1 to each sample to form a G-quadruplex, and then add 10 μL of a HEPES buffer solution containing 2 μM hemin, and react at room temperature for 1 h to form a G-quadruplex-hemorin Vegetarian complex. Finally, add 15 μL of TMB-H to each sample 2 O 2 Solution, G-quadruplex-heme catalyzes TMB color development. After 4 minutes of reaction, 200 μL of 2 M sulfuric acid is added to terminate the catalytic reaction. Characterize the constructed logic gate using ultraviolet-visible ...

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Abstract

The invention discloses a DNA colorimetric logic gate construction method based on metal ion regulation and control of exonuclease III shearing action and belongs to the technical field of molecular computer. Double-stranded DNA containing T-T or C-C mismatched base cannot be sheared by exonuclease III. When there exists Hg<2+> or Ag<+>, 3' end completely matched double-stranded DNA formed by action of T-Hg<2+>-T or C-Ag<+>-C can be sheared by exonuclease III, and released single-stranded DNA hybridizes with a DNA signal probe 1 or DNA signal probe 2, thus leading to formation or disintegration of a G-quadruplex structure. Hereby, Hg<2+> and Ag<+> are used as input signals, and logic operation is carried out through the DNA signal probe so as to construct multiple DNA colorimetric logic gates.

Description

technical field [0001] The invention relates to a method for constructing a DNA colorimetric logic gate based on metal ions regulating the shearing action of exonuclease III, and belongs to the technical field of molecular computers. Background technique [0002] Silicon crystal-based computers perform Boolean logic operations by receiving Boolean input signals and generating corresponding electronic output signals. Similarly, molecular computing includes many bottom-up methods to design biomolecules or chemical molecules, and has become another hot research field of computer technology. Researchers have been working on biological and chemical systems that can perform logical operations at the molecular level. DNA logic devices can be constructed by hybridization reactions between nucleic acid molecules, by toehold-controlled strand displacement reactions, by beacon molecular probes, by aptamer binding, by deoxygenation and enzymatic functional reactions construction, so t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G06F19/20
CPCC12Q1/6839G16B25/00C12Q2537/119
Inventor 邱建丁郜湾梁汝萍
Owner NANCHANG UNIV
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