Establishment of Two-dimensional Gel Electrophoresis for Proteomic of Bursa of Fabricius

To establish a method of two-dimensional electrophoresis(2-DE) in good repetition and high resolution for proteomic analysis of bursa of Fabricius,the bursa of Fabricius from specific pathogen free chickens at different days old were taken to analyses by 2-DE.The first-dimensional electrophoresis was immobilized pH gradients(IPG) strips isoelectric focusing and second-dimensional electrophoresis was vertical electrophoresis of SDS-PAGE.The conditions for sample preparation,rehydration,isoelectric focusing,equilibration,staining and other steps were modified.The results were analyzed with the software PDQuest8.0.1.More than 800 spots on 17 cm IPG strip(pH 5-8) for normal bursa of Fabricius tissue were detected with this method,and the match rate of protein spots was 83.5%.Comparative analysis of bursa of Fabricius at different ages in multiple 2-DE gels revealed 37 differentially expressed protein spots,including 17 up-regulated protein spots,11 down-regulated protein spots,5 new emerging protein spots and 4 disappear protein spots.The method of 2-DE for proteomic of bursa of Fabricius has been established.It would provide a basis for further elucidation of development and immune function of bursa of Fabricius.