本文選題：煙曲霉 + 致病機制�。� 參考：《吉林大學(xué)》2010年博士論文

【摘要】： 本研究以臨床重要病原真菌煙曲霉(Aspergillus fumigatus)為材料,通過連續(xù)傳代的方法,獲得表型和致病力均發(fā)生明顯變化的突變株。對野生株和突變株的胞內(nèi)蛋白質(zhì)、分泌蛋白質(zhì)進行比較蛋白質(zhì)組學(xué)研究,以及野生株的溫度敏感性與致病性關(guān)系的研究,亦進行了不同菌株間胞內(nèi)蛋白質(zhì)的比較。胞內(nèi)蛋白質(zhì)圖譜分析顯示,有42個差異蛋白質(zhì)點,通過質(zhì)譜分析鑒定出9種。其中5種屬于熱休克蛋白家族(heat shock protein,Hsps),分別是Hsp90、Hsp88、Hsp70、HscA和Sti1。其他4種蛋白質(zhì)分別為真核翻譯起始因子EIF 4、核糖體60S亞基P0和2種肌動蛋白VIP 1和Act1。應(yīng)用實時熒光定量分析技術(shù)對Hsp90、Hsp88、Sti1和EIF 4在基因水平的變化進行探討,發(fā)現(xiàn)了3種熱休克蛋白的mRNA水平和蛋白水平的表達一致。分泌蛋白質(zhì)的研究中發(fā)現(xiàn),野生株中有57種表達上調(diào),突變株僅有11種上調(diào)。在不同溫度(25℃和37℃)下,野生株胞內(nèi)蛋白質(zhì)有55個差異點,鑒定出6種:蛋白酶體調(diào)節(jié)亞單位Rpt3、熱休克蛋白(Hsp82和Hsp BiP/Kar2)和3種“假想”蛋白質(zhì)。上述實驗發(fā)現(xiàn)熱休克蛋白可能與致病性和溫度變化有關(guān)。另外,還比較了來自臨床和環(huán)境的6株煙曲霉的致病力差異,并對強毒株和弱毒株的胞內(nèi)蛋白質(zhì)進行了比較,選擇其中49個差異點,鑒定出2種:菌絲體過氧化氫酶(CatB)和ATP合酶-β亞單位(ATP synthase F1,beta subunit)。并得到1種疑似Hsp70同系物。研究結(jié)果表明,熱休克蛋白家族和過氧化氫酶、一些參與蛋白質(zhì)合成、能量合成的蛋白質(zhì)可能與煙曲霉的致病性相關(guān)。
[Abstract]:In this study, we used Aspergillus fumigatusas, an important pathogenic fungus, as the material, and obtained the mutants with obvious phenotype and pathogenicity by the method of continuous passage. The comparative proteomics of intracellular proteins and secretory proteins of wild and mutant strains and the relationship between temperature sensitivity and pathogenicity of wild strains were studied. The intracellular proteins of different strains were also compared. Analysis of intracellular protein map showed that there were 42 differential protein spots and 9 species were identified by mass spectrometry. Five of them belong to the heat shock protein family, Hsp90, Hsp88, Hsp70, HscA and Stil, respectively. The other four proteins were eukaryotic translation initiation factor EIF 4, ribosomal 60S subunit P0 and two actin VIP 1 and Act1, respectively. The changes of Hsp90 Hsp88-Sti1 and EIF _ 4 at gene level were studied by real-time fluorescence quantitative analysis. It was found that the mRNA level and protein level of three kinds of heat shock proteins were consistent. In the study of secreting proteins, 57 species of wild strains were up-regulated, while only 11 of mutant strains were up-regulated. At different temperatures (25 鈩，

本文編號：1938174