期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 402, 期 1, 页码 299-314出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-011-5409-1
关键词
Two-dimensional difference gel electrophoresis; Mass spectrometry; Reactive oxygen species; Genetically modified organisms
资金
- Funda ao de Amparo a Pesquisado Estado de Sao Paulo (FAPESP, Sao Paulo, Brazil)
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq, Brasilia, Brazil)
- Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES, Brasilia, Brazil)
- Financiadora de Estudos e Projetos (FINEP, Brasilia, Brazil)
- Brazilian Synchrotron Light Laboratory - Associacao Brasileira de Tecnologia de Luz Sincrotron - Proteomic Network of the Sao Paulo state
This work reports the evaluation of differentially expressed enzymes and proteins from transgenic and nontransgenic soybean seeds. Analysis of malondialdehyde, ascorbate peroxidase (EC 1.11.1.11), glutathione reductase (EC 1.6.4.2), and catalase (EC 1.11.1.6) revealed higher levels (29.8, 30.6, 71.4, and 35.3%, respectively) in transgenic seeds than in nontransgenic seeds. Separation of soybean seed proteins was done by two-dimensional polyacrylamide gel electrophoresis, and 192 proteins were identified by matrix-assisted laser desorption/ionization (MALDI) quadrupole time-of-flight (QTOF) mass spectrometry (MS) and electrospray ionization (ESI) QTOF MS. Additionally, the enzyme CP4 EPSPS, involved in the genetic modification, was identified by enzymatic digestions using either trypsin or chymotrypsin and ESI-QTOF MS/MS for identification. From the proteins identified, actin fragment, cytosolic glutamine synthetase, glycinin subunit G1, and glycine-rich RNA-binding protein were shown to be differentially expressed after analysis using the two-dimensional difference gel electrophoresis technique, and applying a regulator factor of 1.5 or greater.
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