S-Space College of Agriculture and Life Sciences (농업생명과학대학) Dept. of Food and Animal Biotechnology (식품·동물생명공학부) Theses (Master's Degree_식품·동물생명공학부)
Quantitative detection of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in soybean using two-dimensional gel electrophoresis
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- 서울대학교 대학원
- CP4 5-enolpyruvylshikimate-3-phosphate synthase; Cp4 5-enolpyruvylshikimate-3-phosphate synthase; 이차전기영동; Two-dimensional gel electrophoresis; MALDI-TOF; Maldi-tof; ESI-MS/MS; Esi-ms/ms; 유전자 재조합 식품
- Thesis (master`s)--서울대학교 대학원 :농생명공학부,2004.
- The Round-up Readyⓡ soybean, which has tolerance against glyphosate, an herbicide, is widely
used for soybean processing. Usually, polymerase chain reaction (PCR) is employed to detect GMOs
at DNA level. Immunological methods including western-blotting and enzyme linked immunosorbent
assay (ELISA) are also used to detect GMOs at protein level. Recently, detection of genetically
modified organisms (GMOs) has become more important because of mandatory labeling.
The objective of this research was the detection and quantification of CP4 5-
enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in GM soybean using two-dimensional gel
electrophoresis (2D-PAGE). Purified CP4 EPSPS from cloned E. coli was used as a control to
monitor the change of CP4 EPSPS in GM soybean and soybean curd. CP4 EPSPS was detected in GM
soybean and GM soybean curd by 2D-PAGE and identified by the matrix-assisted laser desorption
ionization time of flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem
mass spectrometry (ESI-MS/MS) through databases. CP4 EPSPS in soybean products was resolved on
2D-PAGE according to its characteristic isoelectric point of 5.1 by isoelectric focusing (IEF)
and molecular mass of 47.5 kDa by SDS-PAGE. Various percentages CP4 EPSPS in soybean were
quantified. The quantitation tests were perfomed on artificial gels with spots varying Gaussian
volumes by 2D software program.
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