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Application of a Small Molecule Fluorescent Probe with Dual Fluorescent Emissions

A fluorescent probe and fluorescence emission technology, which can be used in organic chemistry, luminescent materials, microbial measurement/inspection, etc., and can solve the cumbersome and complicated detection process of dsDNA

Active Publication Date: 2021-06-11
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In view of the above defects or improvement needs of the prior art, the present invention provides an application of a fluorescent probe with dual fluorescence emission. Due to its unique molecular structure, the fluorescent probe molecule exhibits The unique dual fluorescence emission phenomenon was discovered, and this phenomenon was applied to the preparation of specific recognition and detection reagents for double-stranded DNA, thereby solving the cumbersome and complicated technical problems of the prior art detection process of dsDNA

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  • Application of a Small Molecule Fluorescent Probe with Dual Fluorescent Emissions
  • Application of a Small Molecule Fluorescent Probe with Dual Fluorescent Emissions
  • Application of a Small Molecule Fluorescent Probe with Dual Fluorescent Emissions

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preparation example Construction

[0056] (1) Preparation of dsDNA: Dissolve the ssDNA purified by HPLC in ultrapure water, mix two ssDNAs of the same amount, dilute, shake evenly at 37°C and 225rad / min for 5-10min for later use, and make ready-to-use . According to the experimental content, two ssDNA sequences include the following situations: complementary pairing, single nucleotide mutation, and complete mismatch.

[0057] (2) Preparation of TPBT aqueous solution: Dissolve the purified TPBT in dimethyl sulfoxide to prepare 10mmol L -1 The mother liquor is reserved. Take 1 μL TPBT mother solution and add it to 999 μL ultrapure water to prepare 1 10 μmol L -1 TPBT aqueous solution.

[0058] (3) Measure the fluorescence intensity of step (2) with a fluorometer, gradually add dsDNA dropwise, and measure the fluorescence spectrum after each drop of dsDNA.

[0059] In some embodiments, calf thymus DNA (ctDNA) is selected as dsDNA. Switching to other dsDNA also has similar experimental phenomena.

[0060] Fur...

Embodiment 1

[0092] Embodiment 1: the AIE performance of fluorescent probe TPBT

[0093] 1) TPBT in good solvent and poor solvent system

[0094] Measure the fluorescence emission spectrum of probe TPBT (concentration is 10 μ M) under different volume ratios of water (good solvent) and ethanol (poor solvent), the volume ratio of ethanol is successively 0%, 10%, 20%, 30%, 40% %, 50%, 60%, 70%, 80%, 90%. figure 2 is the AIE performance of 10 μM fluorescent probe TPBT in Example 1. Such as figure 2 As shown in (a), as the volume ratio of ethanol increases, the red fluorescence intensity of TPBT centered at 640nm increases gradually, and when the ratio of ethanol reaches 90%, the emission peak intensity increases by 30 times. It shows that TPBT has AIE performance and has a unique fluorescence emission peak at 640nm.

[0095] 2) Interaction between TPBT and polyanionic heparin sodium

[0096] Heparin was gradually added dropwise to the aqueous solution of photosensitizer TPBT with a con...

Embodiment 2

[0101] Example 2: New 540nm fluorescence emission occurs when the probe TPBT interacts with dsDNA

[0102] Gradually add ctDNA dropwise to the aqueous solution with a concentration of 10 μM probe TPBT, and the concentration of ctDNA is 0-20 μg·mL -1 , the excitation wavelength is 450nm, and the fluorescence spectrum is measured. Such as image 3 As indicated, dropwise add 0-10 μg·mL in TPBT -1 The fluorescence emission peak of ctDNA and TPBT is around 640nm; continue to drop ctDNA to 11μg·mL -1 , TPBT has a new fluorescence emission peak at 540nm, and the fluorescence emission peak at 640nm still exists; continue to drop ctDNA to 20μg·mL -1 , the fluorescence emission peak at 540nm was obviously enhanced gradually. The inserted pictures are TPBT and 10 μg·mL -1 and 20μg·mL -1 The fluorescence pictures of ctDNA under ultraviolet light can show that the fluorescence color and intensity of the two concentrations of ctDNA are significantly different.

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Abstract

The invention belongs to the technical field of molecular detection, and more specifically relates to the application of a small molecule fluorescent probe with dual fluorescence emission. When the small-molecule fluorescent probe interacts with double-stranded DNA, double fluorescence emission occurs under the excitation condition of an excitation wavelength of 350-550nm, and the strongest fluorescence emission peaks are 640±5nm and 540±5nm respectively. Due to its unique molecular structure, the fluorescent probe molecule exhibits a unique dual fluorescence emission phenomenon when it interacts with double-stranded DNA. This phenomenon is used to prepare specific recognition reagents for double-stranded DNA. This solves the technical problem of complicated and complicated detection process of dsDNA in the prior art. The fluorescent probe has a structure shown in formula (1): wherein, R is: or n is an integer of 1-12.

Description

technical field [0001] The invention belongs to the technical field of molecular detection, and more specifically relates to the application of a small molecule fluorescent probe with dual fluorescence emission. It specifically relates to a triphenylamine polypyridinium fluorescent probe with aggregation-induced emission (AIE) properties. Due to its unique dual fluorescence emission phenomenon, the fluorescent probe is suitable for specific recognition of double-stranded DNA (double-stranded DNA) , dsDNA), under this premise, it also has the application of detecting single nucleotide polymorphisms (Single nucleotidepolymorphisms, SNP) and dsDNA damage. Background technique [0002] The DNA chain is a double helix structure constructed by two complementary nucleotide sequences, which can carry all the genetic information of organisms. In recent years, due to its importance in genetic diseases, clinical diagnosis and molecular biology, DNA has been studied and analyzed in gre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C07D417/14C09K11/06
CPCC07D417/14C09K11/06C09K2211/1029C09K2211/1051C12Q1/68C12Q2563/107
Inventor 罗亮孟凡玲何珍艳高玉婷
Owner HUAZHONG UNIV OF SCI & TECH
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