Abstract: In order to obtain the best protein extraction methods of maize grain for 2-DE, and establish the best grain protein 2-DE technology system, quality of the proteins extracted by 3 different extraction methods were analyzed, and analysis of 2-DE for comparison in the same condition was done, results showed that the protein extracted by soluble protein extraction method had high purity, moderate concentration and the most abundant protein spots of 2-DE, was most suitable for the study of 2-DE;in order to determine the most appropriate isoelectric focusing conditions, isolated distribution clearer protein spots, use the protein extracted by soluble protein extraction method, protein electrophoresis sample quantity and IEF parameters were optimized, the most suitable pH range of IPG precast gel for maize grain protein was determined, binding protein MALDI-TOF-TOF mass spectrometry analysis, a set of protein 2-DE technique method suitable for maize grain was established, that is, protein extraction use the soluble protein extraction method, isolated distribution use the 24 cm, pH 4-7 IPG dry strip, the proper amount of sample for electrophoresis was 900 μg, IEF procedures:500 V, 1 h, 1 000 V, 1 h, 2 000 V, 1 h, 4 000 V, 2 h, 8 000 V, 4 h, 8 000 V, 80 000 Vhs, 500 V keep.It was laid the foundation for the further study of grain development related proteins.

Key words: Maize, Grain, Proteomics, Two-dimensional electrophoresis

CLC Number: 

• S513.03