Optimization of Proteome Analysis for Wine Yeast Strains

Proteome analysis is a powerful tool for the investigation of yeast in their natural habitats. Two-dimensional gel electrophoresis has been used extensively in proteomics, but reproducibili-ty is notoriously difficult to achieve with this technology. An optimized procedure allowing high reproducibility for the analysis of wine strains of Saccharomyces cerevisiae and Saccharomyces bayanus has been developed and the critical procedures leading to irreproducibility identified. The steps most impacting gel reproducibility are sample preparation (including accurate protein concentration determination), pouring of the gels, staining, and spot detection. Each of these steps has been optimized for the analysis of the proteome of commercial and native isolates of Saccharomyces . Careful attention must also be paid to other steps in the procedure as well.