Synthesis method and application of pore-diameter-adjustable Co-based MOF material with nucleic acid screening function

A synthesis method and technology of nucleic acid molecules, applied in the direction of biochemical equipment and methods, measurement/inspection of microorganisms, etc., can solve the problems that the interaction of biological macromolecules is in its infancy, and achieve easy adjustment, good separation effect, and simple preparation process Effect

Active Publication Date: 2020-05-26
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

MOFs have been used in the fields of small molecule separation, gas storage, catalysis, imaging, drug delivery, etc., but the research on their interaction with biomacromolecules is still in its infancy

Method used

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  • Synthesis method and application of pore-diameter-adjustable Co-based MOF material with nucleic acid screening function
  • Synthesis method and application of pore-diameter-adjustable Co-based MOF material with nucleic acid screening function
  • Synthesis method and application of pore-diameter-adjustable Co-based MOF material with nucleic acid screening function

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Synthesis of MOF ligands

[0047] (1): Add 4-bromo-2-methoxybenzoic acid methyl ester and pinacol diboronic acid ester to the N,N-dimethylformamide solution according to the equivalent ratio of 1:1.5. Heated to 85°C under the same conditions and reacted for 5 hours, the product was separated by column chromatography, and intermediate 1 was obtained after rotary evaporation and drying.

[0048] (2): Add 4-bromo-2-methoxybenzoic acid methyl ester and intermediate 1 to 1,4-dioxane and aqueous solution in an equivalent ratio of 1:1, wherein 1,4-dioxane The volume ratio of water and water is 4:1, heated to 90 degrees under anhydrous and anaerobic conditions for 24 hours, and the product is separated by column chromatography. After rotary evaporation and drying, it is added to 50 equivalents of dichloromethane solution, and the - Add 8 equivalents of boron tribromide solution at 78 degrees, react for 12 hours, then quench with excess water, then add 5 equivalents of 1M sodiu...

Embodiment 2

[0059] Particle size distribution test:

[0060] The dried sample is prepared with a single crystal silicon zero background sample stage, and then the structure is tested on a powder X-ray diffraction instrument.

[0061] Analysis of results:

[0062] figure 1 It is the powder crystal diffraction pattern of Co-based IRMOF-74-II and simulated Co-based IRMOF-74-II in Example 1. It can be seen from the figure that the synthesized Co-based IRMOF-74-II structure conforms to the structure of the matching simulation;

[0063] figure 2 It is the powder crystal diffraction pattern of Co-based IRMOF-74-III and simulated Co-based IRMOF-74-II in Example 1. It can be seen from the figure that the synthesized Co-based IRMOF-74-III structure conforms to the structure of the matching simulation;

[0064] image 3 It is the powder crystal diffraction pattern of Co-based IRMOF-74-IV and simulated Co-based IRMOF-74-IV in Example 1. It can be seen from the figure that the synthesized Co-base...

Embodiment 3

[0071] Selective Adsorption of Nucleic Acids with Different Spatial Size Structures by MOF Materials

[0072] (Step 1) Design nucleic acids with different structures: ssDNA, dsDNA, G4-DNA, ssRNA, G4-RNA, hairpin-RNA. And label these nucleic acid sequences with terminal fluorescent molecules.

[0073] (Step 2) Test the adsorption efficiency of MOF materials to these nucleic acids with different structures

[0074] (Step 2a) Mix 4 μL of annealed DNA / RNA of different structures (10 μM) with 4 μL of Co-IRMOF-74-II, -III, IV (2 mg / mL) in 50 μL of aqueous solution (containing 100 mM KCl and 20 mM KAc, pH 6 .8), react in a mixer at a constant temperature of 37°C for 2 hours.

[0075] (Step 2b) Measure the fluorescence intensity of the supernatant after centrifugation. The absorption efficiency was calculated using the following formula.

[0076]

[0077]

[0078] (FIoriginal is the fluorescence intensity of pure fluorescently labeled DNA / RNA in a buffer solution without MOF ...

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Abstract

The invention relates to a synthesis method and application of a pore-diameter-adjustable Co-based MOF material with a nucleic acid screening function. The synthesis method comprises the following steps of: (1) respectively synthesizing organic ligands II, III and IV of which the chain lengths are progressively increased as shown in the following formula, (2) respectively preparing the same topological structure by selecting metal Co and the organic ligands II, III and IV to obtain MOFs materials of which the pore diameters are progressively increased, and (3) activating the prepared MOFs material, so as to obtain a Co-based MOF material, namely Co-IRMOF-74-II, Co-IRMOF-74-III and Co-IRMOF-74-IV. The synthesized MOF material is suitable for rapid separation of various types of nucleic acidmolecules with secondary structures and has universality, and MOF materials with different pore sizes are selected according to different types of separated nucleic acid structures.

Description

technical field [0001] The present invention relates to the technical field of synthesizing metal-organic framework materials and the technical field of selective separation of nucleic acids with different secondary structures, in particular to a synthesis method and application of a Co-based MOF material with adjustable pore size and nucleic acid screening function. Background technique [0002] Most nucleic acids, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), exist in linear and flexible structures, but some of them can also form various rigid secondary structures, such as double helix, G quadruple strand Body (G4)1-4, hairpin structure, triple helix and i-motif structure. The secondary structure of RNA molecules is critical to their biological functions and cellular regulation. The conformational dynamics of RNAs are the basis of RNA regulatory mechanisms and the origin of their complex functions. RNA molecules are able to switch between different se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G83/00C12Q1/6806
CPCC08G83/008C12Q1/6806C12Q2523/308
Inventor 周翔邓鹤翔彭双别秉霖
Owner WUHAN UNIV
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