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Method for fixing value of standard substance of G2 EPSPS protein solution

A protein solution and standard substance technology, applied in the biological field, can solve the problems of super weeds, environmental pollution, etc., and achieve the effect of ensuring stable preservation, traceability, and accuracy.

Inactive Publication Date: 2018-10-02
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But on the other hand, the glyphosate-resistant pollen of transgenic crops drifts in the environment, which may lead to the spread of glyphosate-resistant genes and produce super weeds, causing environmental pollution

Method used

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  • Method for fixing value of standard substance of G2 EPSPS protein solution
  • Method for fixing value of standard substance of G2 EPSPS protein solution
  • Method for fixing value of standard substance of G2 EPSPS protein solution

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] (1) Characterization of the basic physical and chemical properties of the standard material raw materials:

[0049] (1) Determination of the purity of G2 EPSPS protein by high performance liquid phase exclusion chromatography: take 7 sub-samples from the purified G2 transgenic protein, dilute the sample to 1 mg / mL with water containing 0.1% TFA, and carry out on the machine Purity analysis, typical chromatogram as figure 1 shown. The results are shown in Table 1, and the average purity was 96.6%.

[0050] Table 1 G2 EPSPS protein HPLC purity analysis results (%)

[0051] Analysis times

1

2

3

4

5

6

7

average value

purity

96.6

96.7

96.5

96.7

96.7

96.6

96.7

96.6

[0052] (2) After gel size exclusion chromatography, the purity of G2 EPSPS protein was analyzed by chip electrophoresis, and 5 G2 protein samples were taken in parallel for analysis. The results are as follows: figure 2 As shown, the targe...

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Abstract

The invention discloses a method for fixing the value of a standard substance of a G2 EPSPS protein solution. The method comprises the following steps: accurately testing the content of a pure G2 EPSPS protein product by using an isotope dilution spectrum method and an amino acid analysis method, finally preparing a G2 EPSPS protein solution standard substance by using a gravimetric method-volumetric method, carrying out value measuring verification by using an ELISA (Enzyme Lined Immunosorbent Assay) method, carrying out uniformity and stability inspection, and carrying out preservation stability verification by putting a stabilizer into the standard substance. The G2 EPSPS protein standard substance prepared by using the method is particularly significant for detecting whether the G2 EPSPS protein in a glyphosate tolerant transgenic crop is reasonably expressed or not. The invention provides the method for fixing the value of the standard substance in the G2 EPSPS protein solution, and the method has good reliability, accuracy and traceability.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for determining the value of a G2 EPSPS protein solution standard substance. Background technique [0002] Since 1996, glyphosate-resistant crops have become the most important GM crops. The United States has planted glyphosate-resistant crops on a large scale since 1998, which has greatly promoted the global development of glyphosate-resistant crops and produced huge economic benefits. But on the other hand, the glyphosate-resistant pollen of transgenic crops drifts in the environment, which may lead to the spread of glyphosate-resistant genes and produce super weeds, causing environmental pollution. Therefore, the preparation of G2 EPSPS protein standard substance is very important for detecting the reasonable expression of G2 EPSPS protein in glyphosate-resistant transgenic crops. [0003] Quantitative detection of G2 EPSPS protein in glyphosate-resistant tr...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/68
CPCG01N33/531G01N33/6848G01N2333/47
Inventor 李亮金芜军林敏宛煜嵩刘卫晓武利庆刘刚柳方方
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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