鸡毒支原体蛋白质组双向电泳体系的建立

Objective To establish and optimize two-dimensional gel electrophoresis method for proteomic analysis of Mycoplasma gallisepticum. Methods Improved ultrasound-assisted urea lysis method is used as extraction method for the M. galliseoticum proteome. Immobiline pH gradients isoelectric focusing was used as 1D and vertical SDS-PAGE was used as 2D. Some factors, such as protein extraction method, volume of loading sample, hydration time, isoelectric focusing procedure, SDS-PAGE electrophoresis procedure, protocol for staining the gels, were improved. Results and Conclusion Two-dimensional gel electrophoresis method of M. gallisepticum was developed, providing a clear background, high resolution, large number of protein spots, and good reproducibility. Optimized 2-DE method can be effectively applied in future research of comparative proteomics study of M. gallisepticum.