蛋白质组学技术平台的建立及其初步应用生物化学与分子生物学专业论文.docxVIP

蛋白质组学技术平台的建立及其初步应用是TFIIH十个亚基中的第七个亚基。AMLl．ETO表达后，MATl在 蛋白质组学技术平台的建立及其初步应用 是TFIIH十个亚基中的第七个亚基。AMLl．ETO表达后，MATl在 胞核中的减少与本实验室观察到的细胞周期同时变慢现象可能相关。 上调的HES5(点1021)为Notchl的靶蛋白。由于Notch通路与干 细胞自我更新的维持密切相关，增多的HES5很可能是使携带 AMLl．ETO的干细胞免除枯竭，等待二次打击的主要原因之一。 综上所述，我们采用蛋白质组学技术手段研究了AMLl．ETO的 表达对细胞产生的早期影响，鉴定出了部分差异蛋白质，为阐明 AMLl．ETO在白血病发病过程中的作用提供了若干线索。 关键词：白血病，AMLl．ETO，蛋白质组学，MORGI，MATl，HES5 lII ABSTRACTCONSTRUCTlON ABSTRACT CONSTRUCTlON OF PROTEOMlC TECHNICAL PLATFORaM AND ITS PRIMARY APPLICATION ABSTRACT AMLl-ETO is chimeric protein generated by chromosomal translocation of t(8； 21)，presenting in 40％FAB·M2 type with abnormal karyotype and 10-15％of all AML patients．However,the role and mechanism that AMLl·ETO exerting in the leukemogenesis is still not fully elucidated at present．The dexrelopment of proteomie technology centering on dynamie and comprehensive resealeh on all the proteins expressed by genome may provide large amount of clues for pertinent work． on the basis of the proteomic platform combining 2DE and MALDI—TOF／TOF mass spectrometry,utilizing AMLl一ETO inducible leukemic cell line of U937一A／E 9／14／1 8，we compared the nuclear protein expression pattern with without AMLl-ETo． In three batches of independent experiments．about 1 1 59±46 spots could be detected on each 2D gel utilizing 17cm pH 3-10 non-linear(NL)gel strip followed by silver-staining．Nineteen stably altered spots were excised for mass spectrometry(MS) identification．Among which 17 spots got their protein identity．All the 17 spots only comprised 14 proteins：3 up·regulated and 1 1 down—regulated Different spots with the saule protein name changed similarly．Here，the fact that both CBFJ3(presented on spots 3117，4109)and SFRS9(presented on spots 2235，3227 and 5206)each aligned linearly the gel suggested that each protein undergo some post_translational modification by small charged molecules(such phosphorylation acetylation) leading to obx，iously altered isoeleetric point(pI)and slightly changed molecular weight(MW) Among