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Quantitation of CP4 5-Enolpyruvylshikimate-3-Phosphate Synthase in Soybean by Two-Dimensional Gel Electrophoresis  

KIM YEON-HEE (Department of Food Science and Technology, School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
CHOI SEUNG JUN (Department of Food Science and Technology, School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
LEE HYUN-AH (Department of Food Science and Technology, School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
MOON TAE WHA (Department of Food Science and Technology, School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Publication Information
Journal of Microbiology and Biotechnology / v.16, no.1, 2006 , pp. 25-31 More about this Journal
Abstract
Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.
Keywords
CP4 5-enolpyruvylshikimate-3-phosphate synthase; GMO; soybean; two-dimensional gel electrophoresis; image analysis;
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Times Cited By Web Of Science : 3  (Related Records In Web of Science)
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