JIOMICS

JIOMICS | VOL 5 | ISSUE 2 | DECEMBER 2015 | 1-62
JOURNAL OF INTEGRATED OMICS
Journal of Integrated Omics
A METHODOLOGICAL JOURNAL
HTTP://WWW.JIOMICS.COM
Special Issue: Proceeding Abstracts of the 4 th International Congress on Analytical Proteomics (ICAP 2015)
Comparative proteomic profile between transgenic and non-transgenic
soybean seeds by 2D-PAGE
B. K. de Campos* 1,2 , M. A. Z. Arruda 1,2
1
Spectrometry, Sample Preparation and Mechanization Group –GEPAM, Institute of Chemistry, University of Campinas –UNICAMP, P.O.
Box 6154, Campinas, SP 13083-970, Brazil. 2 National Institute of Science and Technology for Bioanalytics,Institute of Chemistry, University of
Campinas – Unicamp,P.O. Box 6154, Campinas, SP 13083-970, Brazil. *Corresponding author: brunah.campos@hotmail.com
Available Online: 31 December 2015
Abstract
Purpose: Evaluate possible differences in proteomic map between transgenic and non-transgenic soybean seeds varieties (RR 7511 and
8200, respectively) due to change in its genome by the insertion of cp4EPSPS gene, which confers to the soybeans tolerance to glyphosate.
Experimental description: A proteomic approach based on 2D-PAGE separation to identify alterations between T and NT profile was used.
The soybean seeds proteins were extracted using a buffer (50 mmol L -1 de Tris-HCl, 1,5 mmol L -1 KCl, 10 mmol L -1 DTT, 1,0 mmol L -1 PMSF e
0,1 % SDS (m/v)) and its precipitation performed with ammonium acetate 0,1 mol L -1 . The precipitate was solubilized in a specific buffer and
quantified with a 2D Quant Kit (according to the manufacturer's instructions). A total of 500 μg of protein, was applied to immobilized pH
gradient strips (pH range 4–7) and they were rehydrated at room temperature for at least 12 h. Isoelectric focusing was performed for a total of
14,300 Vh. The second-dimension separation was carried out at 25 mA per gel and 100 W on 1-mm 12% (w/v) SDS polyacrylamide gels. After
separation, the gels were stained with colloidal Coomassie brilliant blue. The gels were scanned using an ImageScanner II and the images processed
by ImageMaster 2D Platinum 6.0, which allowed the estimation of the number of spots, the pI values and molecular mass of the proteins.
Results: The proteomic profile of T and NT seeds is different, thereby more than 20% of concentration of total protein was found in T
seeds. Additionally, throughout the 3D evaluation, 14 different spots were found in the T and only 3 in NT seeds, as exemplified in Figure 1.
This indicates that protein synthesis was different between T and NT soybeans. Intensities of the differentials spots were also observed. For a
more accurate assessment, a quantitative analysis by 2D-DIGE will be performed to assess these changes. As future prospects, we will also
quantify and identify these alterations in the proteomic profile and then propose a biochemical pathway.
Conclusions: The insertion of gene cp4EPSPS in soybean may be changing not only their genome but also their proteomic profile, thus
most probably affecting future generations of plant. Regarding to the human food safety, no case has been reported however bioaccessibility
tests need to be performed.
Keywords: Soybean, Organism Genetically Modified, Transgenic, Proteins, Proteomics, Electrophoresis.
Acknowledgements: The authors thank to FAPESP, CAPES and CNPq for financial supporting.
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