Abstract: In order to obtain sheep seminal plasma 2-dimensional gel electrophoresis maps,the total protein number of samples processed by acetone precipitation and combined different loading quantities on 2-DE map were compared. The results showed that seminal plasma total protein was subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and twenty eight protein bands with different molecular weights,ranging from 14.4 to 116 ku,were identified on the gel. When the loading quantity were 0.8,1.0 and 1.2 mg,the total protein spots could be acquired were 207±10, 281±13 and 374±16 proteins were from 20 to 80 ku and IP from 4.0 to 9.0,respectively. With the increasing of loading quantity,the molecular weight of 20 to 80 ku protein spots increased significantly,but the percentage of protein spots in each molecular weight region were more constant. It was concluded that the protein map of sheep seminal plasma proteome could be constructed with the acetone precipitation method and combining with the optimized loading quantities. The results from this study might be used as a plantform for further studies on sheep seminal plasma proteomic.

Key words: sheep; seminal plasma protein; two-dimensional polyacrylamide gel electrophoresis analysis; proteome