不同浓度和梯度的SDS-PAGE胶对双向电泳中蛋白分离的影响

目的：探讨双向电泳中不同浓度和梯度的SDS-PAGE胶对肠道菌蛋白分离效果的影响。方法：制备弗氏2a志贺菌2457T野生株37℃晚期全菌蛋白质样品,进行不同浓度及梯度的SDS-PAGE,研究分离肠道菌蛋白最适宜的SDS-PAGE胶浓度。结果：获得了3个不同浓度（10%、12.5%和15%）的均一胶电泳图谱和3个不同梯度（4%～15%、10%～20%和12%～14%）的电泳图谱,并比较了这些图谱的分离效果;同时,为了分析肠道菌天然表达蛋白的相对分子质量范围,鉴定了8个极端相对分子质量蛋白。结论：对于肠道菌蛋白质的分离来说,12.5%的均一胶或12%～14%的梯度胶较为适宜。

Effect of Concentrations and Gradient of SDS-PAGE Gel on Proteins Separation in Two-Dimensional Gel Electrophoresis

Objective： To study the effect of different systems of homogeneous gels in a series of concentrations and different gradient gels of SDS-PAGE on enterobacterium proteins separation in two-dimensional gel electrophoresis （2-DE）. Methods： The whole cell proteins expressed by Shigella flexneri 2a 2457T wild-type strain in stationary-phase at 37℃ were separated by different systems of homogeneous gels in a series of concentrations and different gradient gels. Results： Three proteome maps of homogeneous gels with different concentrations（10%, 12.5% and 15%） and three proteome maps of different gradient gels（4%-15%, 10%-20% and 12%-14%） were obtained. The effects of protein separation of these maps were compared. To analyze the range of mass weight of expressed proteins of bacteria, a total of 8 proteins with extreme mass weight were identified. Conclusion： For the separation of enterobacterium proteins in 2-DE, 12.5% homogeneous gel and 12%-14% gradient gel are appropriate.