Antimicrobial experimenting method capable of determining antimicrobial protein fragment molecular weight

An experimental method and technology of protein fragments, which are applied in the field of antibacterial experiments that can determine the molecular weight of antibacterial protein fragments, can solve problems such as heavy workload, influence of foreign proteins, and influence on the accuracy of test data, and achieve easy and simple operation Effect

Inactive Publication Date: 2015-06-10
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current antibacterial zone method cannot determine the fragment size of the protein with antibacterial effect, and there are more inconveniences in later experiments. It is necessary to conduct antibacterial tests on proteins o

Method used

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  • Antimicrobial experimenting method capable of determining antimicrobial protein fragment molecular weight
  • Antimicrobial experimenting method capable of determining antimicrobial protein fragment molecular weight
  • Antimicrobial experimenting method capable of determining antimicrobial protein fragment molecular weight

Examples

Experimental program
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Effect test

Embodiment 1

[0065] Example 1 Experiment of antibacterial effect of Kangtaibao on enterotoxigenic Escherichia coli:

[0066] The experimental method of the bacteriostatic effect of Kangtaibao on enterotoxigenic Escherichia coli comprises the following steps:

[0067] Step 1, prepare 12% separating gel and 5% stacking gel according to Table 1, set aside;

[0068] Table 1 Preparation of polyacrylamide gel electrophoresis stacking gel and separating gel

[0069]

[0070] Step 2: After pouring the separating gel and stacking gel respectively, add an appropriate amount of electrode buffer into the electrophoresis tank to ensure that there are no air bubbles in the electrophoresis tank. Then use a micro-injector to add Contipro and standard protein into the corresponding wells. The sample volume is 20 μL, and the empty wells should be filled with 20 μL of loading buffer to ensure that the current is evenly distributed in the gel.

[0071] Step 3. Adjust the voltage so that when the sample...

Embodiment 2

[0078] Example 2 Bacteriostasis experiment of Kangtaibao on chicken Escherichia coli

[0079] The bacteriostatic experiment of Kangtaibao on chicken Escherichia coli includes the following steps:

[0080] Step 1. Prepare 12% separating gel and 5% stacking gel according to Table 1, and after perfusion respectively, add an appropriate amount of electrode buffer solution into the electrophoresis tank to ensure that there are no air bubbles in the electrophoresis tank. Then use a micro-injector to add Contipro and standard protein into the corresponding wells. The sample volume is 20 μL, and the empty wells should be filled with 20 μL of loading buffer to ensure that the current is evenly distributed in the gel.

[0081] Step 2. Adjust the voltage so that when the sample is in the stacking gel, the control voltage is 80V. When the sample enters the separation gel, adjust the voltage to 100V. After electrophoresis, when the bromophenol blue indicator band is close to the bottom o...

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Abstract

The invention provides an antimicrobial experimenting method capable of determining an antimicrobial protein fragment molecular weight, and relates to the technical field of microorganism. After a reagent, separation gel and spacer gel are prepared, protein for antimicrobial experimenting is subjected to protein electrophoresis, an obtained protein electrophoresis film is subjected to decolorization and dyeing, and after the size of a protein molecular weight is observed through imaging, the protein electrophoresis film is used for the antimicrobial experiment. After the protein electrophoresis film is placed in a PBS buffer solution and balanced for 10 minutes, the protein electrophoresis film is placed in an agar culture dish. A bacterium solution in a logarithmic phase is taken out and added into a nutrient agar culture medium at a temperature of 50 DEG C, and the bacterium solution with a volume concentration of one percent is obtained. The bacterium solution is taken out and poured into the culture dish containing the decolorized protein electrophoresis film, the growth situation of thallus in the culture medium is observed, and the size of protein fragment molecular weight which has the good antimicrobial effect is determined. The antimicrobial experimenting method used for determining the size of the antimicrobial protein fragment molecular weight with the antimicrobial effect has the advantages of being easy to operate, accurate, rapid and cheap. The antimicrobial experimenting method has a wide application prospect.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to an antibacterial experimental method capable of determining the molecular weight of antibacterial protein fragments. Background technique [0002] The tests used to determine the effectiveness of antibacterial drugs in inhibiting bacterial growth in vitro are called bacteriostatic tests. Through the antibacterial experiment, the minimum inhibitory concentration of a drug can be determined to evaluate the antibacterial performance of the drug, which is the most basic pharmacodynamic data of antibacterial drugs. The main methods are diffusion method for qualitative determination (such as plaque inhibition test) and dilution method for quantitative determination (such as minimum inhibitory concentration test). Commonly used methods are constant broth dilution method, micro broth dilution method, agar dilution method, disc diffusion method and E test (concentration gradient a...

Claims

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Application Information

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IPC IPC(8): G01N27/447
Inventor 牛明福张婷婷侯颖马丽苹李阳宫强秦翠丽万鹏
Owner HENAN UNIV OF SCI & TECH
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