摘要
目的建立人类巨细胞病毒(HCMV)感染小鼠脑组织前后的比较蛋白质组学双向电泳技术研究体系。方法昆明系小鼠40只,随机分为两组:实验对照组(20只)和病毒感染组(20只),在接种HCMVAD169株30d后,取小鼠脑组织,研磨、超声裂解、抽提蛋白,采用双向凝胶电泳分离,进行染色,Image Master 2D Platinum软件分析、识别差异表达蛋白质,应用基质辅助激光解吸电离飞行时间质谱得到相应肽质量指纹图谱,搜索数据库鉴定部分差异蛋白质点,并对其中部分蛋白质的表达进行免疫印迹的验证。结果从双向电泳图谱上获得蛋白质斑点:实验对照组(1096±82)个,感染HCMV组(1210±101)个,发现显著差异点36个,对其中6个点进行了筛选鉴定;免疫印迹实验结果也较为吻合。结论利用蛋白质组学技术,检测正常小鼠脑组织及HCMV感染的小鼠脑组织的差异蛋白质,为疾病的早期诊断、致病机制及有效药物靶点的发现提供实验和理论依据。
Objective To establish the two-dimensional electrophoresis profiles with high resolution and reproducibility from the brain tissues of mice with human cytomegalovirus（HCMV） infection.Methods Forty Kunming mice were randomized into HCMV infection group（n=20） with HCMVAD169 injection and control group（n=20） with saline injection in the brain.Thirty days after the injections,the brain tissue of the mice were taken and the protein fractions were isolated by two-dimensional gel electrophoresis（2-DE）.Image Master 2D software was used to identify the differentially expressed proteins,and the peptide mass fingerprint（PMF） data were obtained for identification of the differential protein spots via database searching.Western blotting was performed to verify the expressions of some of the differential proteins.Results and Conclusion The 2-D maps of the brain tissues with high Well resolution and reproducibility were obtained.Some of the differentially expressed proteins identified by mass spectrometry（MS） matched their counterparts in the SWISS-2DPAGE database.Western blotting analyses verified the differential expression of the individual proteins.These data can be of value for studying the diagnosis,pathogenesis and effective therapeutic targets of the disease.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2010年第2期341-344,共4页
Journal of Southern Medical University
基金
深圳市科技计划项目(200802051)
