Analysis Of Cipk Proteome Of Arabidopsis Thaliana Magnesium Sensitive Mutant

Using magnesium sensitive Arabidopsis thaliana mutant CIPK3/9/23/26 and wild type Col0 as the research object,we studied the differential expression of proteins between mutant and wild type of Arabidopsis thaliana.In this paper,the removal methods of two high abundance proteins were compared.By means of proteomics,the difference proteins of leaves of Arabidopsis mutant CIPK3/9/23/26 and wild type Col0 were analyzed,and the differential proteins were identified by mass spectrometry.The role of calcium signal sensor protein CIPK protein kinase may serve as a data reference for target protein.Due to the high proportion of Rubisco protein in the leaves,the presence of high abundance proteins in the proteomics study interferes with the presentation of low abundance proteins,interferes with the resolution of bi-directional gel electrophoresis and thus affects the accuracy of protein identification.Therefore,in order to better study the Arabidopsis leaf proteomics and improve the resolution of two-dimensional gel electrophoresis,it is very necessary to eliminate Rubisco protein.First,the extraction of leaf protein from Arabidopsis thaliana was optimized.We used two methods for removal of high-abundance protein Rubisco in this experiment(the PEG pre-separation method combined with TCA/ acetone and PEG pre-separation method combined with SDS/ phenol).It was found that using PEG pre separation method combined with TCA/ acetone protein strip was more clear and the removal effect of high abundance protein was better.The results showed that PEG combined with TCA/ acetone separation process has a good removal effect.The optimized PEG pre-separation method combined with TCA/ acetone extraction method was used to extract the protein of Arabidopsis CIPK 3/9/23/26 mutant and wild type Col0,and then proteins were seperated by two dimensional electrophoresis technique.The differential proteins were analyzed by ImageMaster 6.0.The differential proteins were identified by mass spectrometry,and Some proteins were identified.The results are as follows: 1)There were 30 differential proteins between wild type and mutant type;2)17 differential proteins were identified by mass spectrometry.These include:non-specific lipid-transfer protein,serine / threonine protein kinase,resistant proteins,protease inhibitors,microtubule related proteins,plastid transcription activator 3,plastid blue protein superfamily protein,50 S ribosomal protein L23,ribose isomerase,active protein,translation initiation factor,ATP combination,Protein disulfide-isomerase.