發(fā)布時(shí)間：2018-01-31 06:03

  本文關(guān)鍵詞： 嗜肺軍團(tuán)菌 效應(yīng)蛋白 釀酒酵母 LegS2 線粒體定位　出處：《中山大學(xué)》2011年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：嗜肺軍團(tuán)菌是軍團(tuán)菌病(Legionnaires' disease)的病原,目前已成為胞內(nèi)病原菌-宿主相互作用研究的理想模型。嗜肺軍團(tuán)菌實(shí)現(xiàn)胞內(nèi)增殖的關(guān)鍵在于吞噬泡中的軍團(tuán)菌能通過(guò)IV(B)型分泌系統(tǒng)分泌大量的效應(yīng)蛋白,改變巨噬細(xì)胞的內(nèi)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(guò)抑制巨噬細(xì)胞的蛋白合成和凋亡等,實(shí)現(xiàn)胞內(nèi)的增殖和擴(kuò)散。近年來(lái),多項(xiàng)研究致力于揭示嗜肺軍團(tuán)菌效應(yīng)蛋白的功能,但由于大部分效應(yīng)蛋白功能冗余并且缺乏有效的檢測(cè)手段,目前對(duì)這些效應(yīng)蛋白的功能還知之甚少。 釀酒酵母(Saccharomyces cerevisiae)是具有類(lèi)似許多真核生物中保守的基礎(chǔ)的生化和細(xì)胞生命過(guò)程的一種單細(xì)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(diǎn),使之成為分子遺傳學(xué)和細(xì)胞生物學(xué)等研究的經(jīng)典模型。目前的研究發(fā)現(xiàn),許多效應(yīng)蛋白在酵母胞內(nèi)的表達(dá)都可引起酵母細(xì)胞表型的改變,而這些蛋白的胞內(nèi)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(shù)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(tuán)菌效應(yīng)蛋白相應(yīng)的宿主靶標(biāo)蛋白的研究中提供新的途徑。在本論文中,我們選擇了幾個(gè)軍團(tuán)菌效應(yīng)蛋白Ceg20 (即lpg1137)、LegK1 (即lpg1483)、RalF (即lpg1950)和LegS2 (即lpg2176),通過(guò)相關(guān)的技術(shù)方案來(lái)研究這些蛋白的胞內(nèi)定位和在釀酒酵母細(xì)胞中表達(dá)的表型變化,發(fā)現(xiàn)了這些效應(yīng)蛋白在酵母細(xì)胞中的表達(dá)均能引起酵母細(xì)胞不同程度的生長(zhǎng)缺陷,并發(fā)現(xiàn)了定位于酵母細(xì)胞線粒體的效應(yīng)蛋白LegS2,效應(yīng)蛋白LegS2還能降低酵母細(xì)胞的耗氧量,提示該效應(yīng)蛋白可能通過(guò)改變宿主線粒體的功能而起作用。以上結(jié)果為闡明這些效應(yīng)蛋白在釀酒酵母細(xì)胞中的功能提供了有用的數(shù)據(jù)。另外,本文也對(duì)應(yīng)用酵母多拷貝校正篩選系統(tǒng)來(lái)篩選軍團(tuán)菌效應(yīng)蛋白的酵母靶標(biāo)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(xué)位授予單位】：中山大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R378

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 康曉明,湯忠群,夏錫榮;嗜肺軍團(tuán)菌感染1例報(bào)告[J];解放軍醫(yī)學(xué)雜志;1982年04期

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本文編號(hào)：1478407