本文選題：腦蛋白質組 + 雙向電泳��； 參考：《中國人民解放軍軍醫(yī)進修學院》2006年碩士論文

【摘要】：目的 本研究旨在建立腦蛋白質組研究的技術平臺,獲得可靠的小鼠腦蛋白質組數據庫,比較和評估小鼠不同發(fā)育階段的腦蛋白質,為下一步進行神經系統(tǒng)疾病的蛋白質組學研究奠定基礎。方法 樣品來源于國際腦蛋白質組委員會所提供的胚胎16天、出生后7天及生后60天等3個發(fā)育時期的C57/B16雌性小鼠腦組織。常規(guī)方法提取組織總蛋白,采用固相pH梯度的二維電泳分離和串聯(lián)質譜分析等進行鑒定,使用Mascot軟件搜索UniProt數據庫,鑒定蛋白種類。結果 獲得3個不同發(fā)育時期小鼠腦組織蛋白質表達譜,完成國際腦蛋白質組計劃的預實驗所要求的質量控制,鑒定出3個發(fā)育階段均有較穩(wěn)定表達的4個蛋白點,包括α烯醇酶、磷蛋白質P19及2個肌動蛋白。經進一步評估后鑒定出隨年齡增大而豐度減低的C14orf166同源蛋白、28×10~3熱/酸穩(wěn)定的磷蛋白、40S核糖體蛋白及演變?yōu)榫o鄰雙點的3-巰基丙酮酸硫基轉移酶。其中α烯醇酶及C14orf166同源蛋白曾有文獻報道與神經發(fā)育及組織發(fā)生有密切相關。并通過蛋白免疫印跡技術對不同發(fā)育階段的α烯醇酶進行了驗證。結論 蛋白質組學的方法為腦發(fā)育研究提供了有參考價值的數據,其差異蛋白的發(fā)現(xiàn)可促進對神經發(fā)育機制的了解,并將為后續(xù)啟動神經系統(tǒng)疾病蛋白質組學研究提供參考。
[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 偽 -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, 偽 -enolase and C14orf166 homologous protein have been reported to be closely related to neurogenesis and histogenesis. The 偽 -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.
【學位授予單位】：中國人民解放軍軍醫(yī)進修學院
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R341

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