志贺菌全菌蛋白质双向电泳技术的建立

Establishment of two-dimensional proteome map of Shigella flexneri

目的：建立针对志贺菌全菌蛋白质组学研究的双向电泳及其相关技术体系。方法：采用超声兼尿素-3-[（3-胆酰胺丙基）-乙二胺]-1-丙磺酸（CHAPS）-二硫苏糖醇（DTT）裂解法提取临床分离志贺菌全菌蛋白，利用同相pH梯度等电聚焦对全菌蛋白进行双向电泳，银染后获得的双向电泳图谱进行Image Master 2D Platinum软件分析，并对实验条件进行调整优化。结果：成功构建了高重复性的志贺菌全菌蛋白质双向电泳罔谱，共获得946±37个蛋白质斑点，pI值主要集中在4．94～7．23之间。结论：通过相关条件的调整优化提高了双向电泳的分辨率，为后续志贺菌蛋白质组学研究奠定了基础。

Aim： To establish a two-dimensional electrophoresis（2-DE） and optimize related techniques in the research of proteomic of Shigella flexneri （ S. flexneri ）. Methods ： Sonication-Urea-3- [ （ 3-,cholamidopropyl ） -dimethylammonio]-i-propanesulfonate （CHAPS）-DTT was employed to extract whole cell proteins in cellular of clinical S.flexneri. And immobilized pH gradient（IPG） 2-DE was adopted. The gels were stained with silver and analyzed by Image Master 2D Platinum software. And a series of steps of 2-DE were optimized. Results： Satisfactory and high repetitive 2-DE maps were obtained. 2-DE and image analysis revealed 946 ± 37 protein spots, and pl value was mainly localized among 4.94 to 7.23. Conclusion： With optimization of the experimental conditions, 2-DE of S. flexneri has been successfully established with high sharpness of separation, which stands as a valuable resource for proteomics research of S.flexneri.