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A method for in situ gel preparation for protease zymography

An in-situ gel and protease technology, applied in the field of soil zymography, can solve the problem that there is no clear high-quality in-situ gel preparation technology, and achieve the effects of sensitive coloring and accurate experimental results.

Active Publication Date: 2021-03-02
HOHAI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Current attempts to localize and quantify protease enzyme activity in soil using protease zymography do not have a well-defined high-quality in situ gel preparation technique or associated parameters

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Preparation of in situ gels (unincubated)

[0033] (1) Dosing: Add 3.5g of agarose, 0.2g of gelatin, 0.5g of β-cyclodextrin and 0.5ml of PBS buffer solution into 250ml of deionized water, and stir evenly.

[0034] (2) Glue: Put the prepared solution in a high-pressure steam sterilizer, and heat it to 70°C under the condition of 0.5mpa, and the heating time lasts for about 10 minutes. Take out until the solution is clear.

[0035] (3) Condensation molding: prepare a glass chamber of 25cm×10cm, the thickness of the glass chamber is 1mm, and the glass chamber must be detachable for the preparation of the gel. Control the temperature of the prepared gel at 70°C and cast it in a glass chamber for cooling and molding (it is basically shaped when it cools to room temperature).

[0036] (4) Staining: Remove the cooled gel. Refer to the ninhydrin method to place the treated gel in the staining solution for 10 minutes.

[0037] The dyeing solution is prepared by mi...

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Abstract

The invention relates to an in-situ gel preparation method for protease zymography and belongs to the technical field of soil zymography. The object of the present invention is to provide a kind of in-situ gel technique that is prepared as experimental raw material with agarose, gelatin, β-cyclodextrin and PBS (phosphate buffered saline solution), can be used for the mensuration of protease distribution in rhizosphere soil, It can also be widely used as experimental material for protease zymography. This in situ gel preparation technology for protease zymography fills the lack of related gel preparation technology. By preparing high-quality in situ gels to incubate the experimental soil, draw a zymogram, and provide a comprehensive overview of the enzymes in the soil. Active calculations are provided for convenience.

Description

technical field [0001] The invention relates to an in-situ gel preparation method for protease zymography and belongs to the technical field of soil zymography. Background technique [0002] With the development and progress of soil zymography, relevant scholars have become more novel in exploring the spatial distribution of microbial activities in soil. Since zymography is an in situ method that does not require disruption of soil structure, it allows for a more realistic picture of enzyme activity. In future research on soil microbes or rhizosphere effects, soil zymography will be more widely popularized and used. [0003] According to existing studies, the level of soil enzyme activity is related to the activity of soil microorganisms, which is of great reference research value in reflecting the status of microbial activity. However, the rhizosphere soil of plants is strongly affected by the accumulation of root exudates, and soil microorganisms are mostly active in this...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08J3/075C08L5/12C08L5/16C08L89/00C12Q1/37
CPCC08J3/075C08J2305/12C08J2405/16C08J2489/00C12Q1/37
Inventor 钱进刘寅陆卞和赵媛媛钱心缘沈俊炜朱鑫泽
Owner HOHAI UNIV
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