Image analysis of two-dimensional gel electrophoresis for comparative proteomics of transgenic and non-transgenic soybean seeds.
Considering the importance of bidimensional electrophoresis and image analysis in comparative proteomics, the parameters that influence the analysis of protein expression of transgenic and non-transgenic soybean seeds were evaluated. The loaded mass of the proteins (150-500 microg), the pH separation range (3-10 or 4-7), and manual/automatic image editing were evaluated. Additionally, after optimizing the conditions, histograms and matchings were obtained in order to accurately analyze the variations (90%) in protein expression. From this, 10 proteins displayed significant differences in expression, and eight of them were characterized and identified by mass spectrometry.
Brandão
A R
AR
Spectrometry, Sample Preparation and Mechanization Group-GEPAM, Institute of Chemistry, University of Campinas-UNICAMP, Campinas, SP, Brazil.
Barbosa
H S
HS
Arruda
M A Z
MA
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted / Q000379:methods; D030821:Plants, Genetically Modified; D040901:Proteomics / Q000379:methods*; D012639:Seeds / Q000737:chemistry; D030262:Soybean Proteins / Q000737:chemistry*; D013025:Glycine max / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D030262:Soybean Proteins