Electroluminescence logic gate adopting adenosine monophosphate and adenosine deaminase as excimers

A technology of adenosine monophosphate and adenosine deaminase, applied in the field of electroluminescent logic gates

Inactive Publication Date: 2014-04-09
LINYI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Obviously, if it is possible to communicate through electrical signal

Method used

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  • Electroluminescence logic gate adopting adenosine monophosphate and adenosine deaminase as excimers
  • Electroluminescence logic gate adopting adenosine monophosphate and adenosine deaminase as excimers
  • Electroluminescence logic gate adopting adenosine monophosphate and adenosine deaminase as excimers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Embodiment 1 Treatment of multi-walled carbon nanotubes and preparation of multi-walled carbon nanotubes modified electrodes

[0012] 2g of multi-walled carbon nanotubes was crushed in 36% HCl for 6 hours, washed with water and filtered to neutrality, and then dissolved in 2.2mol / L HNO 3 After the supernatant was crushed for 1 hour, it was cultured in an oven at 90°C for one night, then it was crushed for 1 hour at room temperature, and washed with water until neutral. Take 1 mg of the treated multi-walled carbon nanotubes and dissolve them in 100 mL of water containing 1 mg of DMF, and then ultra-pulverize for 1 h until a black suspension is formed.

[0013] After rinsing, the oxidized glassy carbon electrode was immersed in 10mmol / L ethylenediamine containing 5mmol / LEDC and 8mmol / LNHS and slightly shaken for 5h. After washing again, the electrode was transferred to the suspension of multi-walled carbon nanotubes containing 5mmol / L EDC and 8mmol / L NHS, and the multi-w...

Embodiment 2

[0014] Example 2 Immobilization of single-stranded DNA on the surface of multi-walled carbon nanotube modified electrode

[0015] Take a certain amount of Ru(bpy) 2 (dcbpy)-S 1 ( figure 1 ) solution was mixed with 2.0mL0.1mol / L PBS buffer solution (pH7.0, 0.1mol / LNaCl), so that the concentration of ssDNA in the solution was 1×10 -6 mol / L, immerse the multi-walled carbon nanotube modified electrode in the above solution for 30 minutes with slight shaking. After fixing the electrode, use 0.2mol / L BR buffer solution (0.5% SDS, 0.04mol / LH 3 BO 3 , 0.04mol / L H 3 PO 4 , 0.04mol / L HAC, pH7.0) and secondary water were washed separately.

[0016] The improved operation takes a certain amount of Ru(bpy) 2 (dcbpy)-S 2 , S 1 and Fc-S 3 Mix in 2.0mL0.1mol / L PBS buffer solution (pH7.0, 0.1mol / L NaCl) so that the concentration of ssDNA in the solution is 1×10 -6 mol / L, after the hybridization is complete, add AMP to perform the same operation as above, which can make Ru(bpy) 2 (...

Embodiment 3

[0017] Example 3 Electrochemiluminescence detection

[0018] This experiment was carried out on the MPI-E electrochemiluminescence analysis system. Three-electrode system: with Ru(bpy) 2 (dcbpy)-ssDNA-modified multi-walled carbon nanotube modified electrode was used as the working electrode, a platinum wire electrode was used as the counter electrode, and Ag / AgCI (saturated KCI) was used as the reference electrode. The buffer solution is 2.0mL containing Ru(bpy) 2 (dcbpy)-ssDNA 0.1mol / LPBS buffer solution (pH7.0, 0.1mol / LNaCl), using cyclic voltammetry scanning (0.3-1.3V), the high voltage of the photomultiplier tube is 800V.

[0019] In the buffer solution, the concentration is 1×10 -6 mol / L of S 1 , S 2 and S 3 The reaction was stirred at a constant temperature of 37° C. for 30 minutes. 10 μL of 1 mmol / L AMP was added to the buffer solution, stirred and reacted at a constant temperature of 37° C. for 30 minutes, and then detected by electrochemiluminescence. After th...

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Abstract

The invention relates to electrochemiluminescence DNA logic gate constructed by adopting adenosine monophosphate and adenosine deaminase as excimers based on a specific binding effect of a aptamer and a ligand thereof, and a deamination effect of adenosine deaminase, wherein adenosine monophosphate aptamer-containing single-stranded DNA and two single-stranded DNA modified with a luminescence agent and a quenching agent are subjected to hybridization to form the DNA logic gate. According to the present invention, after adenosine monophosphate and the aptamer thereof are subjected to specific binding, the luminescence agent-modified single-stranded DNA is released, and is captured by the carbon nanotube-modified electrode so as to enhance the electrochemiluminescence signal; adenosine deaminase is added to catalyze deamination of adenosine monophosphate so as to destruct the binding effect of the adenosine monophosphate and the aptamer, and the aptamer and the luminescence agent-modified single-stranded DNA form the hybrid so as to separate from the electrode and reduce the electrochemiluminescence signal; and through the improvement of the logic gate structure and the introduction of the quenching agent, the background interference is reduced, and the detection sensitivity is improved.

Description

technical field [0001] The invention relates to the construction of a DNA logic gate, in particular to an electroluminescent logic gate using adenosine monophosphate and adenosine deaminase as exciters. Background technique [0002] Ribozymes and nucleic acid aptamers are nucleic acid structures that are widely concerned now, and they can be DNA, RNA, or a hybrid of DNA and RNA. At present, nucleic acids and ribozymes have been widely used in the development of DNA logic gates, but the application of nucleic acid aptamers has not been reported. The input and output signals of current DNA logic gates or computers are mainly realized through molecular biology operations or fluorescent readers. Obviously, if it is possible to communicate by electrical signals, it is possible to significantly improve the transmission efficiency. The nucleic acid aptamers of biomacromolecules and cells modified with electrochemiluminescent molecules and quenching groups are immobilized on the e...

Claims

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Application Information

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IPC IPC(8): G01N21/76
Inventor 郭英姝李雪梅李伟
Owner LINYI UNIVERSITY
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