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Preparation method for in-situ gel applied to protease zymography

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

Active Publication Date: 2020-06-09
HOHAI UNIV
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  • Summary
  • 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 a preparation method for in-situ gel applied to protease zymography, belonging to the technical field of soil zymography. The invention aims to provide a technology for preparing the in-situ gel by taking agarose, gelatin, beta-cyclodextrine and a PBS (phosphate buffered solution) as experimental raw materials. The in-situ gel can be used for determining protease distribution in rhizosphere soil and can also be widely applied as an experimental material for protease zymography. According to the preparation technology for the in-situ gel applied to protease zymography,a blank in related gel preparation technologies in the prior art is filled in; and experimental soil is incubated by preparing the high-quality in-situ gel, a zymogram is drawn, and convenience is provided for calculation of the activity of enzymes in the soil.

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