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神经蛋白质组学双向电泳技术体系的建立

Establishment of a two-dimensional electrophoresis technologic platform on neuroproteomics

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【作者】 刘师莲秦延江张旭华靖永胜徐广润王晓明王伟

【Author】 LIU Shi-lian1, QIN Yan-jiang2, ZHANG Xu-hua1, JING Yong-sheng1, XU Guang-run2, WANG Xiao-ming1,WANG Wei1 (1.Experimental Center of Molecular Biology, School of Medicine, Shandong University, Jinan 250012, Shandong, China;2.Department of Neurology, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China)

【机构】 山东大学医学院分子生物学实验中心山东大学齐鲁医院神经内科山东大学医学院分子生物学实验中心 山东济南250012山东济南250012山东济南250012

【摘要】 目的:建立神经蛋白质组学双向电泳技术体系。方法:利用双向电泳技术分别以人脑脊液、人和鼠脑组织为研究对象,通过离心等手段提取蛋白,行双向电泳,硝酸银染色,进行图谱分析。结果:从双向电泳图谱上获得蛋白质斑点:正常人脑脊液359个,多发性硬化人脑脊液397个,正常人脑组织480个,正常鼠脑组织524个。经与SWISS-2DPAGEDATA比对,发现差异蛋白点。结论:从蛋白质水平探讨多发性硬化症、脑出血等疾病的发病机制,对寻找有效的药物靶点具有重要意义,同时为神经系统其它疾病的研究提供了切实可行的神经蛋白质组学研究技术体系。

【Abstract】 Objective: To establish a two-dimensional electrophoresis technology platform on neuroproteomics. Methods: Cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) and brain tissues of patients with intracerebral hemorrhage (ICH) were taken to establish a technical system by two-dimensional electrophoresis. In sample preparation, overdrive centrifugation and sedimentation were used. During isoelectric focusing(IEF), different length of immobilized pH strips and different composition of rehydrating buffer were applied. And during sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the same 12%-concentration of PAG was used. Silver nitrate was applied to stain proteins. Results: Different 2-dimentional maps were got. There were 359 spots detected in CSF of human control, 397 in CSF of patients with MS, 480 in brain tissue of human, and 524 in brain tissue of rat. Conclusion: The technology of proteome is applied in the study of CSF and brain tissue. It is helpful in revealing the pathogenesis of nervous system through displaying some special proteins, and offers some drug targets for the therapy of diseases.

【基金】 山东省科技厅立项课题(96k984)。
  • 【文献出处】 山东大学学报(医学版) ,Journal of Shandong Univenity(Health Scicnes) , 编辑部邮箱 ,2005年05期
  • 【分类号】R341
  • 【被引频次】11
  • 【下载频次】369
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