摘要
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.
出处
《郑州大学学报(医学版)》
CAS
北大核心
2007年第2期316-318,共3页
Journal of Zhengzhou University(Medical Sciences)
关键词
志贺菌
蛋白质组学
双向电泳
Shigellaflexneri
proteomics
two-dimensional electrophoresis