2-DE Technical Platform Of Chorispora Bungeana Differential Proteomics

Since the completion of genome sequences of human, attention has been focused on the post-genomic era of which the development of proteomics is the symbol. Although many techniques have been used successfully for proteomic analysis,2-DE is still the essential technique currently for separation of complex protein mixtures. Sample preparation is crucial in two-dimensional gel electrophoresis(2-DE), also being the prerequisite in the proteomics analysis. It is difficult to get satisfying result of 2-DE because of the serious interference of pigment, phenols, quinones, and other plenty of secondary metabolites from plant tissues. Therefore, construction of a high quality technical platform of 2-DE is crucial for analysis of the whole-cell proteins.In this experiment ,an optimized protocol was developed, paying special attention to the whole-cell protein analyses of Chorispora bungeana based on 2-DE.The optimization included the thiourea-acetone/TCA-methyl alcohol-phenol method, Ready Strip IPG of 5-8 pH range,12% gel concentration,and Coomassie Blue G-250 as staining reagent. At last, a proper SDS-PAGE gel was obtained. We expect that this work will have some contributions in discovering molecular mechanisms at the level of proteins and establishing the network of molecular regulation.