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Preparation method and application of fluorescently-labeled aptamer metal organic framework probe

A metal-organic framework and fluorescent labeling technology, which is used in the preparation and application of mercury ion intracellular imaging nanoprobes, can solve problems such as easy degradation, achieve simple methods, promote development, and increase immobilization capacity

Pending Publication Date: 2022-02-15
CHONGQING MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, since the aptamer itself belongs to the oligonucleotide chain, it is easy to degrade, and its own negative charge limits its entry into the cell to play a role

Method used

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  • Preparation method and application of fluorescently-labeled aptamer metal organic framework probe
  • Preparation method and application of fluorescently-labeled aptamer metal organic framework probe
  • Preparation method and application of fluorescently-labeled aptamer metal organic framework probe

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Embodiment 1

[0039] Step 1. Disperse 48 mg of terephthalic acid and 67.2 mg of zirconium tetrachloride each in 12 mL of DMF solution to obtain a clear solution. After mixing the two, 16 mL of DMF solution was added, and then 2 mL of acetic acid was added while stirring. After heating in an oil bath at 90°C for 12 hours, the Zr-MOFs solution was obtained, which was taken out and cooled to room temperature. Centrifuge at 10,000rpm / min for 5min, wash with absolute ethanol for 3 times, and vacuum dry overnight to obtain Zr-MOFs nanomaterials in the form of white solid;

[0040]Step 2. Separately disperse T20 DNA and A10 DNA in Tris-HCl, and then mix them at a molar concentration ratio of 1:1.5. Heat the mixture in a water bath at 95°C for 10 minutes, then place it at room temperature for 1 hour to fully hybridize the T20 / A10 double strands;

[0041] Step 3. Mix 100 μg / mL Zr-MOFs with 100 nM double-stranded DNA, and incubate at room temperature for 1 hour to obtain the mixture;

[0042] Step...

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Abstract

The invention relates to a preparation method of a fluorescently-labeled aptamer metal organic framework probe and application of the fluorescently-labeled aptamer metal organic framework probe in the field of mercury ion intracellular imaging. AMCA-labeled T20 is used as an ion recognition element, and mercury ions are recognized through formation of a T-Hg<2+>-T structure. Dabcyl-labeled A10 is hybridized with the T20, so that a fluorescence signal is changed from nothing to the existence. Meanwhile, a Zr-MOFs nano material serves as a carrier for hybrid double chains to enter cells. The characteristic of high specific surface area of the Zr-MOFs nano material is utilized, and more DNA double strands are adsorbed through combination of zirconium unsaturated sites and a DNA phosphoric acid skeleton, while the DNA double strands are protected from being degraded by an external complex environment. The probe has the advantages of high sensitivity, strong specificity, and rapid and convenient detection.

Description

Technical field: [0001] The invention belongs to the field of preparation and application of mercury ion intracellular imaging nanoprobes, and in particular relates to a preparation method of a fluorescence-labeled aptamer metal organic framework probe and its application in mercury ion intracellular imaging. Background technique: [0002] With the growth of the national economy, environmental pollution is becoming more and more serious all over the world. This situation has attracted the attention of relevant departments. Among them, the harm of heavy metal pollution to residents' health and public safety has become increasingly apparent. Mercury is a heavy metal element with accumulation and multi-affinity. Excessive accumulation in the body can cause serious damage to the human immune system, blood system, nervous system and reproductive system, and even be carcinogenic. my country has included mercury pollution as one of the key prevention and control contents of nation...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806G01N33/53C12Q1/02C09K11/06C08G83/00G01N21/64
CPCC12Q1/6806G01N33/5308G01N33/5008C09K11/06C08G83/008G01N21/6458G01N21/6428C09K2211/188G01N2021/6432C12Q2525/205C12Q2563/107
Inventor 何俊琳郑姝婷于超
Owner CHONGQING MEDICAL UNIVERSITY
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