本文關鍵詞： 嗜肺軍團菌 效應蛋白 釀酒酵母 LegS2 線粒體定位　出處：《中山大學》2011年碩士論文　論文類型：學位論文

【摘要】：嗜肺軍團菌是軍團菌病(Legionnaires' disease)的病原,目前已成為胞內病原菌-宿主相互作用研究的理想模型。嗜肺軍團菌實現(xiàn)胞內增殖的關鍵在于吞噬泡中的軍團菌能通過IV(B)型分泌系統(tǒng)分泌大量的效應蛋白,改變巨噬細胞的內吞途徑,阻止溶酶體與吞噬泡的融合,使自己免于被消化,并通過抑制巨噬細胞的蛋白合成和凋亡等,實現(xiàn)胞內的增殖和擴散。近年來,多項研究致力于揭示嗜肺軍團菌效應蛋白的功能,但由于大部分效應蛋白功能冗余并且缺乏有效的檢測手段,目前對這些效應蛋白的功能還知之甚少。 釀酒酵母(Saccharomyces cerevisiae)是具有類似許多真核生物中保守的基礎的生化和細胞生命過程的一種單細胞生物,其易于培養(yǎng)、遺傳背景清楚和基因操作方便等特點,使之成為分子遺傳學和細胞生物學等研究的經(jīng)典模型。目前的研究發(fā)現(xiàn),許多效應蛋白在酵母胞內的表達都可引起酵母細胞表型的改變,而這些蛋白的胞內定位也為了解其功能提供有益的線索。另外,作為研究蛋白-蛋白相互作用技術方案之一的釀酒酵母多拷貝校正篩選系統(tǒng)也可以在篩選和鑒定軍團菌效應蛋白相應的宿主靶標蛋白的研究中提供新的途徑。在本論文中,我們選擇了幾個軍團菌效應蛋白Ceg20 (即lpg1137)、LegK1 (即lpg1483)、RalF (即lpg1950)和LegS2 (即lpg2176),通過相關的技術方案來研究這些蛋白的胞內定位和在釀酒酵母細胞中表達的表型變化,發(fā)現(xiàn)了這些效應蛋白在酵母細胞中的表達均能引起酵母細胞不同程度的生長缺陷,并發(fā)現(xiàn)了定位于酵母細胞線粒體的效應蛋白LegS2,效應蛋白LegS2還能降低酵母細胞的耗氧量,提示該效應蛋白可能通過改變宿主線粒體的功能而起作用。以上結果為闡明這些效應蛋白在釀酒酵母細胞中的功能提供了有用的數(shù)據(jù)。另外,本文也對應用酵母多拷貝校正篩選系統(tǒng)來篩選軍團菌效應蛋白的酵母靶標蛋白作了初步的嘗試。
[Abstract]:Legionnaires pneumophila is the pathogen of Legionnaive 'disease. It has become an ideal model for the study of intracellular pathogen-host interaction. The key to the intracellular proliferation of Legionella pneumophila lies in the ability of phagocytic Legionella to pass through IVB. The secretory system secretes a large number of effector proteins. To change the endocytosis pathway of macrophages, to prevent the fusion of lysosome and phagocytic vesicles, to prevent themselves from being digested, and to realize the proliferation and diffusion of macrophages by inhibiting the protein synthesis and apoptosis of macrophages. Many studies have been devoted to revealing the function of Legionella pneumophila effector proteins, but because most of them are redundant and lack effective detection methods, little is known about the functions of these effector proteins. Saccharomyces cerevisiae is a single-celled organism that has a conserved basis of biochemical and cellular life processes similar to that of many eukaryotes. It is easy to culture, genetic background is clear and easy to operate, making it a classical model of molecular genetics and cell biology. The expression of many effector proteins in yeast cells can cause phenotypic changes in yeast cells, and the intracellular localization of these proteins also provides useful clues for understanding their functions. As one of the techniques to study protein-protein interaction, the Saccharomyces cerevisiae multi-copy correction screening system can also provide a new way to screen and identify the corresponding host target proteins of Legionella. In this paper. We have selected several Legionella effector protein Ceg20 (lpg1137) and LegK1 (lpg1483). RalF (i.e. lpg1950) and LegS2 (lpg2176). The intracellular localization and phenotypic changes of these proteins in Saccharomyces cerevisiae cells were studied by relevant technical schemes. It was found that the expression of these effector proteins in yeast cells could cause the growth defects of yeast cells to varying degrees, and the effector protein LegS2 located in yeast cell mitochondria was also found. The effector protein LegS2 can also reduce the oxygen consumption of yeast cells. These results provide useful data for elucidating the function of these effector proteins in Saccharomyces cerevisiae cells. The yeast target protein of Legionella effect protein was screened by yeast multiple copy correction screening system.
【學位授予單位】：中山大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R378

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相關期刊論文 前1條

1 康曉明,湯忠群,夏錫榮;嗜肺軍團菌感染1例報告[J];解放軍醫(yī)學雜志;1982年04期

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本文編號：1478407