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Method for acquiring massspectrogram of standard protein spot of differential expression induced by uranium of common pokeweed

A technique of differential expression and uranium contamination, which is applied in the field of acquisition of standard protein spot mass spectrograms to achieve high sensitivity

Inactive Publication Date: 2011-02-16
HUNAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to detect uranium pollution safely and accurately. It provides a method for obtaining the standard protein spot mass spectrogram of Pokeweed uranium-induced differential expression for uranium pollution detection, and also provides a method using American Method for detection of uranium contamination by spot mass spectrometry of differentially expressed standard proteins induced by pokeweed uranium

Method used

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  • Method for acquiring massspectrogram of standard protein spot of differential expression induced by uranium of common pokeweed
  • Method for acquiring massspectrogram of standard protein spot of differential expression induced by uranium of common pokeweed
  • Method for acquiring massspectrogram of standard protein spot of differential expression induced by uranium of common pokeweed

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Experimental program
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Effect test

Embodiment 1

[0030] Production of standards for differentially expressed proteins in the detection of uranium contamination:

[0031] (1) Collect pokeweed roots in the uranium tailings area and pokeweed in the non-polluted control area, and store pokeweed roots in the uranium tailings area and the non-polluted control area in liquid nitrogen at -196°C;

[0032] (2) Preparing whole protein by saturated phenol-ammonium acetate / methanol precipitation method;

[0033] (3) The 2D electrophoresis method combining isoelectric focusing and polyacrylamide was used to display the whole protein, and stained with Coomassie brilliant blue method;

[0034] (4) The stained gel block was scanned by UMAX Power Look 2100 gel image scanner at 600dpi, and the obtained image was analyzed by PDQuest software;

[0035] (5) Screen the differentially expressed protein spots, perform in-gel enzymatic hydrolysis (Trypsin, 20 hours), extract enzymatic hydrolyzed peptides, and mass spectrometry analysis on the differ...

Embodiment 2

[0037] Detection method of pokeweed uranium pollution in plants to be tested

[0038] Collect the roots of Pokeweed in the environment to be tested and the pokeweed in the non-polluted control area, and freeze the pokeweed in the environment to be tested and the root tissue of the pokeweed in the non-polluted control area in liquid nitrogen at -196 ° C; other steps are the same as in Example 1. Finally, the similarity comparison between the obtained differentially expressed protein spot mass spectrograms and the identified five uranium-induced differentially expressed standard protein spot mass spectrograms was carried out, so as to accurately identify whether the environment was polluted by uranium.

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Abstract

The invention discloses a differential expression protein standard used for detecting uranium pollution, which is a standard protein spot spectrogram of differential expression under the induction of five pokeberries, and the uranium pollution in the environment can be accurately detected by taking the standard protein spot spectrogram of the differential expression under the five vegetal pokeberries as a standard for detecting the uranium pollution; in addition, the safe detection can be carried out away from polluted environments; and the invention has high sensitivity and solves the problem that the uranium pollution in ambient environments at the pollution area is difficult to be detected accurately.

Description

technical field [0001] The invention relates to the technical field of detecting environmental pollution by molecular biology methods, in particular to a method for obtaining and using a standard protein spot mass spectrogram of differentially expressed Pokeweed uranium-induced differential expression for uranium pollution detection. Background technique [0002] The traditional uranium pollution and the detection of radioactive substances in the environment are usually detected by radioactive detectors, and the probes of α, β, χ, γ multifunctional ray detectors are used to detect the radiation dose. The disadvantages of this traditional detection method are: (1 ) Although it can detect a certain dose of radioactive source, it is difficult to distinguish what kind of radioactive substance it is; (2) its sensitivity has certain limitations; threaten. [0003] American pokeweed (Phytolacca americana L.pokeweed), also known as vertical pokeweed, ten core pokeweed. A large num...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/62
Inventor 高利臣肖璐冯涛
Owner HUNAN UNIV OF SCI & TECH
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