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A kind of green fluorescent protein and layered composite metal hydroxide composite luminescent ultra-thin film and preparation method thereof

A technology of green fluorescent protein and layered compounding, applied in the direction of fluorescence/phosphorescence, chemical instruments and methods, luminescent materials, etc., to achieve the effect of improving utilization rate, good stability and repeatability

Active Publication Date: 2016-10-05
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the introduction of fluorescent proteins into the interlayers of layered materials LDHs

Method used

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  • A kind of green fluorescent protein and layered composite metal hydroxide composite luminescent ultra-thin film and preparation method thereof
  • A kind of green fluorescent protein and layered composite metal hydroxide composite luminescent ultra-thin film and preparation method thereof
  • A kind of green fluorescent protein and layered composite metal hydroxide composite luminescent ultra-thin film and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0030] 1. Prepare LDHs by fast nucleation crystallization isolation method, the specific method is to weigh 15.385gMg(NO 3 ) 2 ·6H 2 O and 11.254g Al(NO 3 ) 3 9H 2 O dissolved in 80ml to remove CO 2 Deionized water, denoted as A solution, Mg 2+ / Al 3+ The molar ratio is 2:1; Weigh 7.2g NaOH and dissolve it in 80ml to remove CO 2 Deionized water, denoted as B solution; A and B two kinds of solutions were added to the total back-mixed liquid membrane reactor at the same time, the slit width between the reactor rotor and the stator was adjusted to 2mm, the working voltage was 140V, and the rotor speed was 5000rpm. The obtained mixed slurry was transferred to a reaction kettle, and after crystallization at 110°C for 24 hours, it was fully washed with deionized water, and the fourth centrifugal liquid was taken as the magnesium-aluminum LDHs colloidal solution. The centrifuged substrate was dried at 60°C for 18 hours to obtain dry magnesium-aluminum LDHs.

[0031]2. Take 1...

Embodiment 2

[0038] 1. Prepare LDHs by hydrothermal method, the specific method is to weigh 15.703g Ni(NO 3 ) 2 ·6H 2 O and 10.129gAl(NO 3 ) 3 9H 2 O dissolved in 80ml to remove CO 2 Ni in deionized water 2+ / Al 3+ The molar ratio is 2:1; weigh 16.8g of hexamethylenetetramine HMT and dissolve it in 120ml to remove CO 2 Deionized water; pour the two solutions into a four-neck flask, mix while stirring, and pass nitrogen gas for 1 hour at the same time, then heat and stir in a water bath at 80°C for 3 days; use to remove CO 2 Centrifuge and wash with deionized water until the pH reaches 6.5-7.0, weigh 5g of the centrifugal substrate (nickel-aluminum LDHs) and dissolve it in 100mL to remove CO 2 Deionized water, nitrogen gas for 30 minutes, put in an oven at 120°C for 12 hours, take it out and centrifuge at a speed of 2000rmp for 30 minutes, and take the supernatant as the nickel-aluminum LDHs colloidal solution.

[0039] 2. With embodiment 1;

[0040] 3. With embodiment 1;

[0041...

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Abstract

The invention discloses a novel bio-inorganic composite luminescent material, namely, a composite luminescent ultrathin film of a green fluorescent protein and a layered composite metal hydroxide, and a preparation method thereof. The preparation method includes following steps: (1) preparing a layered composite metal hydroxide colloidal solution through a quick-nucleation crystallization isolation method or a hydrothermal synthetic method; (2) preparing a fluorescent protein solution; (3) treating a substrate with concentrated sulfuric acid and hydrogen peroxide and washing the substrate respectively with alcohol and water; and finally (4) preparing the composite luminescent ultrathin film of the green fluorescent protein and the layered composite metal hydroxide through a layer-by-layer self-assembling method. The composite luminescent ultrathin film is excellent in pH-responding stability and repeatability, can be used for reversible detection of biological in-vivo pH environments, can increase the utilization rate of the fluorescent protein and provides a basis of application researching and researching of other performances of the fluorescent protein on a bio-sensor.

Description

technical field [0001] The invention belongs to the technical field of bio-inorganic composite material preparation, and specifically relates to a method for preparing a green fluorescent protein / LDHs composite luminescent ultra-thin film by a layer-by-layer self-assembly method, and evaluates its performance. Background technique [0002] Green Fluorescent Protein (GFP) was discovered in 1962. Because it can emit stable fluorescence, has no toxicity, is not species-specific, and is easy to express in cells, it has been widely used as a marker in life. science field. When the GFP gene is fused with the target protein gene and expressed in vivo, the target protein still maintains normal activity, and GFP can maintain its fluorescence, so the location, movement and other activities of the target protein can be tracked. As the most effective living cell markers, fluorescent proteins can be used to study the regulation of gene expression, the interaction of biological macromole...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C09K11/06G01N21/64
Inventor 陆军张平段雪
Owner BEIJING UNIV OF CHEM TECH
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