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
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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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