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United States Department of Agriculture

Agricultural Research Service

Title: Analysis of Soybean Seed Development by Difference Gel Electrophoresis

Authors
item Miernyk, Jan
item Johnston, Mark
item Herman, Eliot
item Mooney, Brian - UNIVERSITY OF MISSOURI

Submitted to: American Society of Plant Biologists Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: March 21, 2005
Publication Date: N/A

Technical Abstract: Embryos were harvested at specific stages (T1-T8) of soybean (Glycine max L. cv. Jack) seed development. Total protein fractions were isolated from seeds at each stage, and the components separated by 2-dimensional electrophoresis. Sequential sets of proteins were labeled with CyDye3 and CyDye5 for difference gel electrophoresis (DIGE) analysis. When these gels were subject to image analysis, proteins present at a constant level appeared as yellow spots, proteins that increased in abundance appeared green, and proteins that decreased in abundance appeared red. Selected proteins were subjected to gel tryptic digestion, and peptide mass fingerprint analysis by MALDI-TOF mass spectrometry. Preparative liquid-phase isoelectric focusing and chromatography with cross-linked concanavalin-A were used to reduce the level of storage proteins in the input samples for 2-D separations, allowing analysis of less-abundant protein spots. Our goal is to prepare a catalog of protein changes during oilseed development as the basis for analysis of the mechanisms that regulate protein remodeling in plant cells.

Last Modified: 10/21/2014
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