Light emitting system of ribonuclease and inhibitor sieving of ribonuclease, preparation method and application of light emitting system

A technique for screening ribonucleases and inhibitors, applied in biochemical equipment and methods, hydrolytic enzymes, fluorescence/phosphorescence, etc., can solve problems such as no relevant reports on gold nanoclusters

Pending Publication Date: 2019-05-21
CHINA THREE GORGES UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

[0004] So far, gold nanoclusters prepared with RNA as a template and their applications have not been reported

Method used

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  • Light emitting system of ribonuclease and inhibitor sieving of ribonuclease, preparation method and application of light emitting system
  • Light emitting system of ribonuclease and inhibitor sieving of ribonuclease, preparation method and application of light emitting system
  • Light emitting system of ribonuclease and inhibitor sieving of ribonuclease, preparation method and application of light emitting system

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Embodiment Construction

[0042] The following experiments and examples serve to further illustrate but not limit the present invention.

[0043] (1) Preparation of gold nanoclusters using RNA as a template

[0044] Add 50 μL of chloroauric acid (HAuCl 4 , 1 mM) and 400 μL ribonucleic acid (RNA, 0.16 mg / mL), mix evenly, after 10 minutes in a water bath at 37°C, slowly add 500 μL freshly prepared ascorbic acid (AA, 1 mM), and stir vigorously for 4 hours. After 4 hours of reduction, 50 μL of mercaptoundecanoic acid (MUA, 1 mM) was added to the mixed solution, and the stirring was continued for 30 minutes. The obtained system was centrifuged at a speed of 2000 rpm for 20 minutes to remove free gold atoms in the solution, and a clear and transparent light pink solution was obtained, which was gold nanoclusters. The photos under 310 nm ultraviolet light irradiation were as follows: figure 1 , to measure the absorption spectrum and fluorescence emission spectrum as figure 2 , the luminous intensity is I ...

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Abstract

The invention relates to a light emitting system of ribonuclease and inhibitor sieving of ribonuclease, a preparation method and application of the light emitting system. Gold nanometer clusters are based on good biocompatibility and emit red light, and are prepared from chloroauric acid (HAuCl4) under mild conditions, with ribonucleic acid (RNA) and mercaptoundecanoic acid being a microenvironment and with ascorbic acid being a mild reducing agent. The bright red fluorescent light is emitted, and the wavelength of the emitted light is about 630 nm. In the system, if a ribonucleic acid shear enzyme (such as RNase A) exists, the RNA structure is destroyed, the gold nanometer clusters cannot be formed, or the light-emitting intensity is low, and therefore the aim of quantitatively and qualitatively determining the activity of the ribonuclease is realized. The system can be further used for sieving the ribonuclease inhibitor. The preparation condition of the gold nanometer clusters is that a water bath is conducted at the temperature of 30-40 DEG C, and the volume ratio of the chloroauric acid to the RNA to the ascorbic acid to the mercaptoundecanoic acid is 0.8-1.2:6-10:8-15:0.8-1.2.Through the light-emitting intensity of the light emitting system, the ribonuclease can be quantitatively and qualitatively detected, wherein the quantitative detection range is 0-2.5*10-4 U / mL.

Description

technical field [0001] The invention relates to a photochemical method for specific detection of ribonuclease (such as RNase A) and its inhibitor screening, which belongs to the field of biomacromolecule enzyme activity detection and enzyme inhibitor screening. Background technique [0002] Gold nanoclusters (AuNCs) are new nanoluminescent materials composed of dozens to hundreds of gold atoms. In recent years, gold nanoclusters have been favored by many researchers because of their unique physical and chemical properties. Sensing and other fields have shown advantages such as good water solubility, low toxicity, biocompatibility, and large stocks displacement, and have broad application prospects. [0003] In the early days, the preparation of gold nanoclusters was realized under the protection or assistance of thiol compounds, polyethyleneimine, polyamide-amine, etc., but the gold nanoclusters obtained by these methods emit blue fluorescence, and tissue penetration in biol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/22C12Q1/48G01N21/64
Inventor 杨昌英杜金娅刘含龚玲俐陆爽
Owner CHINA THREE GORGES UNIV
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