脑发育不同阶段蛋白质组学及血清脑红蛋白含量的研究

发布时间：2018-07-10 14:26

本文选题：脑蛋白质组 + 双向电泳　；参考：《中国人民解放军军医进修学院》2006年硕士论文

【摘要】：目的本研究旨在建立脑蛋白质组研究的技术平台,获得可靠的小鼠脑蛋白质组数据库,比较和评估小鼠不同发育阶段的脑蛋白质,为下一步进行神经系统疾病的蛋白质组学研究奠定基础。方法样品来源于国际脑蛋白质组委员会所提供的胚胎16天、出生后7天及生后60天等3个发育时期的C57/B16雌性小鼠脑组织。常规方法提取组织总蛋白,采用固相pH梯度的二维电泳分离和串联质谱分析等进行鉴定,使用Mascot软件搜索UniProt数据库,鉴定蛋白种类。结果获得3个不同发育时期小鼠脑组织蛋白质表达谱,完成国际脑蛋白质组计划的预实验所要求的质量控制,鉴定出3个发育阶段均有较稳定表达的4个蛋白点,包括α烯醇酶、磷蛋白质P19及2个肌动蛋白。经进一步评估后鉴定出随年龄增大而丰度减低的C14orf166同源蛋白、28×10~3热/酸稳定的磷蛋白、40S核糖体蛋白及演变为紧邻双点的3-巯基丙酮酸硫基转移酶。其中α烯醇酶及C14orf166同源蛋白曾有文献报道与神经发育及组织发生有密切相关。并通过蛋白免疫印迹技术对不同发育阶段的α烯醇酶进行了验证。结论蛋白质组学的方法为脑发育研究提供了有参考价值的数据,其差异蛋白的发现可促进对神经发育机制的了解,并将为后续启动神经系统疾病蛋白质组学研究提供参考。
[Abstract]:Objective to establish a technical platform for the study of brain proteome, to obtain a reliable proteome database of mouse brain, and to compare and evaluate the brain proteins of mice at different stages of development. To lay a foundation for proteomics research of nervous system diseases. Methods the samples were collected from the brain tissues of C57 / B16 female mice during the developmental period of 16 days, 7 days after birth and 60 days after birth, which were provided by the International Committee on brain Proteomics. Tissue total protein was extracted by routine method and identified by solid-phase pH gradient two-dimensional electrophoresis and tandem mass spectrometry. Mascot software was used to search the UniProt database to identify the protein types. Results three protein expression profiles of mouse brain tissue were obtained at different developmental stages. The quality control required for the pre-experiment of the international brain proteome program was completed. Four stable protein spots were identified in all the three developmental stages. It includes 伪 -enolase, phosphorous protein P19 and two actin. After further evaluation, the C14orf166 homologous protein, 28 脳 10 ~ (-3) pyruvate stable phosphoroprotein 40S ribosomal protein and developed into a 3-mercaptopyruvate thiotransferase, was identified with increasing age. Among them, 伪 -enolase and C14orf166 homologous protein have been reported to be closely related to neurogenesis and histogenesis. The 伪 -enolase at different developmental stages was verified by Western blot. Conclusion the method of proteomics provides valuable data for the study of brain development. The discovery of differential proteins can promote the understanding of neurodevelopmental mechanism and provide a reference for the subsequent study of proteomics of nervous system diseases.
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R341

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