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Molecular and immunological characterization of the major outer membrane proteins of Brucella
Authors:Axel Cloeckaert,Jean-Michel Verger,Maggy Grayon,Nieves Vizcaí  no
Affiliation:Department of General and Marine Microbiology, Lundberg Laboratory, University of Göteborg, Medicinaregatan 9C, 413 90 Göteborg, Sweden
Abstract:
Abstract Saccharomyces cerevisiae exponentially growing in basic or 0.7 M NaCl medium were isotopically labelled with 35S-methionine, followed by protein separation and quantification by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) combined with computerised image analysis. The electrophoretic separation resolved about 650 proteins of which 13 displayed significant and at least 2-fold changes in rate of synthesis during saline growth. By sequencing of 2D-PAGE resolved proteins, one of the 8 induced spot, p42.9/5.5, was shown to correspond to the full length (containing the N-terminal extension) product of the GPD 1 gene encoding the cytoplasmic glycerol 3-phosphate dehydrogenase. The expression of the TDH 3 gene, glyceraldehyde 3-phosphate dehydrogenase, and the ENO 2 gene, enolase, decreased during growth in NaCl medium, declines hypothesised to have an impact on the flux to glycerol.
Keywords:Saccharomyces cerevisiae    NaCl stress    Protein synthesis    2D-PAGE    Protein sequencing    GPD1    Glycolytic enzymes
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