Proteome Comparative Analysis Of The Seedling Of PhyAphyB Mutant And Columbia Wild-type-4 Of Arabidopsis Thaliana

Plant functional genomics is developing very fast in this century. However, the information of gene express on the transcription level still can hardly confirm the gene's function. Because the direct execute factor of life is protein other than the gene. Moreover, in the process of transcription and translation, the protein varies in different organism, different tissue even different period. So, proteomics witch was a leading technology for large-scale analysis of protein expression and function has become a major field of functional geomics. The technology of proteomics required to separate large numbers of proteins, to identify them, and to study their modifications are by no means straight forward. Currently the only separation technology which provides highly purified proteins separated in a simple parallel process is 2-Delectrophoresis. In proteome projects, which aim to identify and characterize all proteins expressed by an organism or tissue, the identification of proteins plays a central role.The proteins 2-D maps of Arabidopsis wild type (Col-4), red light receptor double mutant (phyAphyB) were obtained by optimizing crucial factors and procedures such as sample treatment, electrophoresis parameter, and gel concentration. The results revealed that adapting lyses solution to treating sample, choosing moderate amount of sample, using pre-cast immobilized linear pH gradient pH 3~10 24cm dry strips for the first dimensional electrophoresis, 12% separation gel were used to the SDS-PAGE electrophoresis. Fifty-five protein spots, which were differentially expressed in the phyAphyB and col-4, were identified by the method: MALDI-TOF-TOF-MS peptide fingerprint analysis of the protein spots and protein database searching. Thirty-nine of them were identified successfully.In order to find out the new factor which operate in both red light pass way and blue light pass way, we analyzed the proteins expression of Arabidopsis Col-4, cry1-304 and phyB grown in white light, blue light and red light for 7-8 days by SDS electrophoreses in this research. Compared the two mutants with Col-4, we find several dissimilitude in each light situation.