Commentaires
Diaporama
Plan
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Proteomics analyses of the wheat kernel proteins
(Proteomics evaluations of ITMI lines)
  • Branlard G.,
  • INRA
  • Clermont-Ferrand
  • France
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Overview
  • 1  The amphiphilic proteins of ITMI progeny
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Wheat proteomics
  • Proteome : all the proteins expressed in a cell (an organite or a tissue) at a given time
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The ITMI population
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Methods
  • 1-     Protein extraction :
  • Tx114  buffer                    (Amphiphilic)
  • Sodium Phosphate buffer (Albumins + Globulins)
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Methods
  • 1-    Protein extraction : Tx114 (amphiphilic), Sodium Phosphate (Albumins + Globulins)
  • 2-      2DE : IPG x SDS-PAGE,
      •  IPG 6-11  for Amphiphilic; IPG 4-7 for Albumins + Globulins
  • Coomassie and Silver staining
  • 3-6 replicates per sample
  • 3-       Scanning and Image analysis (Melanie, Genebio):
  • gel matching,  spot detection,  spot analyses
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Methods
  • 1-    Protein extraction : Tx114 (amphiphilic), Sodium Phosphate (Albumins + Globulins)
  • 2-      2DE : IPG x SDS-PAGE,
      •  IPG 6-11  for Amphiphilic; IPG 4-7 for Albumins + Globulins
  • Coomassie and Silver staining
  • 3-6 replicates per sample
  • 3-       Scanning and Image analysis (Melanie, Genebio):
  • gel matching,  spot detection,  spot analyses
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Mass spectrometry
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Methods
  • 1-    Protein extraction : Tx114 (amphiphilic), Sodium Phosphate (Albumins + Globulins)
  • 2-     2DE : IPG x SDS-PAGE,
      •  IPG 6-11  for Amphiphilic; IPG 4-7 for Albumins + Globulins
  • Coomassie and Silver staining
  • 3-6 replicates per sample
  • 3-    Scanning and Image analysis (Melanie, Genebio):
  • gel matching,  spot detection,  spot analyses
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Data Base interrogations
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RESULTS
  • 1- The amphiphilic proteins of ITMI progeny
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Mapping on Ch1D of loci coding for amphiphilic proteins
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Amphiphilic proteins of Synthetic and Opata
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Identification of the proteins in the puroindoline zone
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Identification of the amphiphilic proteins
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RESULTS
  • 1  The amphiphilic proteins of ITMI progeny
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 Albumins and Globulins specific to Opata and to Synthetic
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 Mapping genes coding for Albumins and Globulins
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 Identification of some Albumins and Globulins
using MALDI-TOF and data bases
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RESULTS
  • 1  The amphiphilic proteins of ITMI progeny
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"Regulation of the quantitative expression..."
  •   Regulation of the quantitative expression of the different loci
    •         interactions between  chromosomes (homologous and homeologous)
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Gluten proteomics :
  • Effect of heat stress on wheat proteome at kernel maturity on cv Thésée
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Projects
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Projects
  • 1-  The European proteomic project of Healthgrain
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Conclusion
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Acknowledgements
  • AMIOUR Nardjis
  • BANCEL Emmanuelle
  • DUMUR Jérome
  • LEROY Philippe
  • MAJOUL Thouraya
  • MERLINO Marielle
  • NADAUD Isabelle