空腸彎曲菌膜融合蛋白CmeA、外膜蛋白CmeC的表達(dá)、純化與結(jié)晶篩選
發(fā)布時(shí)間：2019-03-19 17:00
【摘要】：空腸彎曲菌(Campylobacter jejuni)是最常見的導(dǎo)致腸炎的細(xì)菌,它也可通過神經(jīng)節(jié)苷脂模擬機(jī)制導(dǎo)致周圍神經(jīng)系統(tǒng)自免疫紊亂的格林巴利綜合征,對人類健康造成危害。臨床經(jīng)驗(yàn)表明,空腸彎曲菌在人體腸道內(nèi)的生存和繁殖過程中,對四環(huán)素、紅霉素和氯霉素敏感,對青霉素、大靈堿、喹啉酮和頭孢霉素表現(xiàn)出很強(qiáng)的耐受性。通過對其感染過程及抗性機(jī)制的了解,有針對性地設(shè)計(jì)藥物與抗體,從而減少空腸彎曲菌對人類健康的危害,成為近年來的一個(gè)較重要的課題。CmeABC系統(tǒng)是空腸彎曲菌中的多重耐藥外排系統(tǒng),在對抗生素以及膽鹽的抗性中起著主要作用。CmeABC系統(tǒng)包括三種蛋白：細(xì)胞膜融合蛋白CmeA,細(xì)胞內(nèi)膜蛋白CmeB和細(xì)胞外膜蛋白CmeC其中CmeA蛋白主要功能為連接CmeB和CmeC,幫助形成外排通道。CmeC蛋白主要是作為通道,將抗生素和膽鹽等從細(xì)菌體內(nèi)運(yùn)到體外。外源性的抗體、藥物分子可以通過作用于外膜蛋白CmeC影響到CmeABC系統(tǒng)的功能,所以它是一個(gè)潛在的藥物作用靶點(diǎn)。本課題對空腸彎曲菌膜融合蛋白CmeA、外膜蛋白CmeC蛋白進(jìn)行表達(dá)、純化和結(jié)晶篩選,為以后解析CmeA, CmeC蛋白的結(jié)構(gòu)、針對性地設(shè)計(jì)藥物、治療相關(guān)疾病打下基礎(chǔ)。并可借此深入理解此類蛋白的結(jié)構(gòu)與功能特征,理解CmeABC系統(tǒng)在空腸彎曲菌生存、增殖和感染中的作用。
[Abstract]:Campylobacter jeejunii is the most common bacteria that leads to enteritis, and it can also cause the peripheral nervous system to self-immune disorder of the Guillain-Barre syndrome through the ganglionic fat simulation mechanism, which is harmful to human health. The clinical experience shows that the Campylobacter jejuni is sensitive to tetracycline, erythromycin and chloramphenicol in the survival and reproduction of human intestinal tract. In order to reduce the harm of Campylobacter jejuni to human health, it has become a more important topic in recent years. The CmeC system is a multi-drug-resistant efflux system in Campylobacter jejuni, and plays a major role in the resistance to antibiotics and bile salts. The CmeC system consists of three proteins: the cell membrane fusion protein CmeA, the cell membrane protein CmeB and the extracellular membrane protein CmeC, wherein the main function of the CmeC protein is to connect CmeB and CmeC to help form an outer discharge channel. The CmeC protein is mainly used as a channel, and antibiotics and bile salts and the like are transported from the bacteria body to the outside. Exogenous antibodies, drug molecules, can influence the function of the CmeC system by acting on the outer membrane protein CmeC, so it is a potential drug action target. The expression, purification and crystallization of CmeC protein of Campylobacter jejuni membrane fusion protein CmeA and outer membrane protein CmeC were selected, and the structure of CmeA and CmeC protein was analyzed in order to lay the foundation for the treatment of related diseases. The structure and functional characteristics of such proteins can be further understood, and the role of the CmeC system in the survival, proliferation and infection of Campylobacter jejuni is understood.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R378
本文編號(hào)：2443718
[Abstract]:Campylobacter jeejunii is the most common bacteria that leads to enteritis, and it can also cause the peripheral nervous system to self-immune disorder of the Guillain-Barre syndrome through the ganglionic fat simulation mechanism, which is harmful to human health. The clinical experience shows that the Campylobacter jejuni is sensitive to tetracycline, erythromycin and chloramphenicol in the survival and reproduction of human intestinal tract. In order to reduce the harm of Campylobacter jejuni to human health, it has become a more important topic in recent years. The CmeC system is a multi-drug-resistant efflux system in Campylobacter jejuni, and plays a major role in the resistance to antibiotics and bile salts. The CmeC system consists of three proteins: the cell membrane fusion protein CmeA, the cell membrane protein CmeB and the extracellular membrane protein CmeC, wherein the main function of the CmeC protein is to connect CmeB and CmeC to help form an outer discharge channel. The CmeC protein is mainly used as a channel, and antibiotics and bile salts and the like are transported from the bacteria body to the outside. Exogenous antibodies, drug molecules, can influence the function of the CmeC system by acting on the outer membrane protein CmeC, so it is a potential drug action target. The expression, purification and crystallization of CmeC protein of Campylobacter jejuni membrane fusion protein CmeA and outer membrane protein CmeC were selected, and the structure of CmeA and CmeC protein was analyzed in order to lay the foundation for the treatment of related diseases. The structure and functional characteristics of such proteins can be further understood, and the role of the CmeC system in the survival, proliferation and infection of Campylobacter jejuni is understood.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R378
【共引文獻(xiàn)】
相關(guān)博士學(xué)位論文 前1條
1 趙麗娜;基層醫(yī)療機(jī)構(gòu)大腸埃希菌及肺炎克雷伯菌氟喹諾酮耐藥分子流行及遺傳特征研究[D];浙江大學(xué);2015年
，本文編號(hào)：2443718
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