绵羊卵泡液蛋白质组2-DE图谱的构建及初步分析

The Mapping and Preliminary Analysis of Ovine Follicular Fluid Proteome 2-DE

为了获得绵羊卵泡液蛋白质组表达图谱,采用2-Dclean-upkit法和热的SDS直接裂解法制备卵泡液蛋白质样品,进行双向凝胶电泳,运用PDQuest8.0对图谱进行初步分析,最后与血浆蛋白质表达图谱做初步对比。结果显示:热的SDS直接裂解法处理的蛋白质样品图谱蛋白质斑点较多,更能真实反映卵泡液蛋白质组的全貌;卵泡液蛋白质图谱与血浆蛋白质图谱匹配率达75.62%。研究表明,热的SDS直接裂解法处理蛋白质样品效果较好;卵泡液蛋白质成分与血浆蛋白质成分比较相似,可以用处理血浆样品的方法来优化处理卵泡液蛋白质样品。

In order to obtain maps of ovine follicular fluid proteome 2-DE, the 2-D clean-up kit method and hot-SDS lysis method were applied to prepare follicular fluid sample, and the samples were subjected to two-dimensional gel electrophoresis. The 2-DE maps of ovine follicular fluid were analyzed through PDQuest8.0, and then were compared with ovine plasma proteome 2-DE maps. The result showed that the map of the sample prepared through hot-SDS lysis method has more protein sports and could reflect the real profile of the follicular fluid proteome better. The follicular fluid proteomic 2-DE map was similar to the plasma proteomic map, and the matching rate reached 75.62%. It was concluded that hot-SDS lysis method was a better way to prepare follicular fluid sample, and the protein composition in follicular fluid was similar to that in plasma. So we can optimize the method that prepare follicular fluid sample according to the method to prepare plasma sample.