Quantitative detection method for transgene protein CP4-EPSPS in plant

A CP4-EPSPS, quantitative detection method technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to apply transgenic protein detection, limit the application range, reduce the quantitative accuracy based on immunology, etc., and achieve the reduction of biological safety Hidden dangers, recovery rate, satisfactory stability, and high sensitivity

Active Publication Date: 2015-07-08
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods require the use of different antibodies according to different target proteins, which brings great challenges to the wide application of the method
And because there are many factors in the process of processing crops into food, the above method cannot be applied to the detection of genetically modified proteins in processed foods, which limits its application range
Additionally, both non-specific binding and cross-contamination can reduce the accuracy of immunology-based quantification

Method used

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  • Quantitative detection method for transgene protein CP4-EPSPS in plant
  • Quantitative detection method for transgene protein CP4-EPSPS in plant
  • Quantitative detection method for transgene protein CP4-EPSPS in plant

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Embodiment Construction

[0021] Embodiments of the present invention are described below with reference to the drawings. Elements and features described in one drawing or one embodiment of the present invention may be combined with elements and features shown in one or more other drawings or embodiments. It should be noted that representation and description of components or processes that are not relevant to the present invention and known to those of ordinary skill in the art are omitted from the drawings and descriptions for the purpose of clarity.

[0022] The present invention will be further described below in conjunction with the accompanying drawings.

[0023] 1 Materials and methods

[0024] 1.1 Reagents (Table 1)

[0025] Table 1 Reagent list

[0026] Reagent name

Specification

production company

Synthetic peptide ITGLLEGEDVINTGK

95.52%

Beijing Saibaisheng Gene Technology Co., Ltd.

Synthetic peptide SFMFGGLASGETR

97.27%

Beijing Saibaisheng ...

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PUM

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Abstract

The invention relates to a quantitative detection method for transgene protein CP4-EPSPS in a plant. The quantitative detection method comprises: extracting protein from a plant to obtain a crude protein extraction solution; and carrying out digestion on the crude protein extraction solution, adopting three 18O-labeled CP4-EPSPS specific peptide segments as internal standards, adopting UPLC / ESI-QQQ MS detection, and carrying out quantification on CP4-EPSPS in a peptide segment level, wherein the three specific peptide segment sequences are ITGLLEGEDVINTGK, SFMFGGLASGETR and LAGGEDVADLR. According to the present invention, the method has high sensitivity, and absolute CP4-EPSPS quantification can be achieved through combination of an isotope dilution method and an MRM strategy.

Description

technical field [0001] The invention relates to the application of proteomics technology, in particular to a method for quantitative detection of transgenic protein CP4-EPSPS in plants by applying isotope dilution method and multiple reaction monitoring (MRM) strategy. Background technique [0002] The CP4-EPSPS gene has been widely used in transgenic crops and plants. Currently, 61% of GM crops / plants comprising 30 different plants are widely grown in 12 countries worldwide. As the number and varieties of genetically modified crops / plants are increasing year by year in many countries, the impact of genetically modified crops / plants on human health and the environment has attracted more and more attention. Therefore, quantitative techniques for detecting transgenic proteins in transgenic crops / plants have gradually become one of the research hotspots. [0003] In recent years, research on the quantitative methodology of transgenic proteins in crops / plants has become one of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
Inventor 邓玉林张玫
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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