Two-dimensional gel electrophoresis model for proteomic analysis of duck plague virus infected duck embryo fibroblasts.

To establish and optimize two-dimensional gel electrophoresis(2-DE) models for proteomic analysis of duck plague virus infecting duck embryo fibroblasts.Various protein extraction methods,Sample loading buffer strip range of IPG,sampling volume of protein,staining method and so on were compared and analyzed.Results:Protein extraction by acetone and reducing agent of 30 mmol/L DTT were easy to be focalized;Loading sampling buffer of mixturation with pH3-10 carrier ampholytes and pH5-8 carrier ampholytes at a 2∶1 ratio with the 17 cm IPG strip of pH 5-8 could improve the resolution.Protein sample of 0.8 mg was fit for Coomassie brilliant blue R-250 staining,while 1.5 mg was too high.It was found that 1 253 protein spots were obtained in 17 cm pH 5-8 gel strip by silver staining while around 388 were obtained by coomassie brilliant blue staining.The spots of 348 and 331 protein were obtained in 17 cm pH 5-8 gel strip each different time in the sample with coomassie stains,and the mean matching rate achieves to 88%.The findings showed that the better repetition,resolution and stability of 2-DE models will be a better method to supply further study of DPV proteomic,and served as an important method for discovering new proteins.