绵羊精浆蛋白质组2-DE图谱的构建及初步分析

The Construction and Preliminary Analysis of Sheep Seminal Plasma Proteomic 2-DE Map

本研究比较了丙酮沉淀法制备蛋白质不同上样量对绵羊精浆蛋白质二维电泳图谱的影响。结果显示,精浆总蛋白经SDS-PAGE电泳,得到分子质量在14.4~116 ku的28条蛋白质条带。二维电泳图经PDQuest 8.0分析,上样量为0.8、1.0、1.2 mg时检测出的蛋白质点分别为207±10、281±13和374±16个,分子质量基本分布在20~80 ku、等电点为4~9的区域内,随着上样量的增加,分子质量在20~80 ku的蛋白质点明显增多,但每个分子质量区间的蛋白质点所占的比率较为恒定。研究结果表明,采丙酮沉淀制备精浆蛋白结合合适的上样量能够建立绵羊精浆全蛋白质图谱,为进一步研究绵羊精浆蛋白质组学奠定基础。

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.