Abstract:Rhizopus chinensis CCTCCM201021 is an important filamentous fungus,which was isolated from Daqu,a traditional leaven in the production of Chinese liquor. In order to understand this fungal deeply,the conditions of two-dimensional electrophoresis(2-DE) for proteome analysis of R. chinensis were investigated and optimized. Intracellular proteins of Rhizopus chinensis was extracted and separated for 2-DE. Different sample preparation methods,parameters of the gel,sample loading quantity and methods were investigated to optimize 2-DE maps of R. chinensis. Ultimately disrupt cell in liquid nitrogen and mechanical agitation,precipitate proteins with TCA/acetone,passively rehydrate proteins with 24 cm pH 4~7 linear strips were selected. The sample loading quantity for each strip was 1 200 μg,and modified colloidal coomassie brilliant blue G-250 was applied to stain gels. Finally,2D-gel map with no obvious vertical and horizontal stripes,low background,high resolution and good reproducibility was obtained. Approximately 800 protein spots were isolated from the 2-DE protein map after analyzed by PD-Quest software.
