本文關(guān)鍵詞：甲型副傷寒沙門(mén)氏菌外膜蛋白的表達(dá)及其免疫保護(hù)性研究　出處：《南昌大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 甲型副傷寒沙門(mén)氏菌 外膜蛋白 克隆 過(guò)量表達(dá) 體內(nèi)攻毒 免疫保護(hù)性

【摘要】： 傷寒是一種急性全身性感染，每年全球傷寒(包括副傷寒)患者約2200萬(wàn)，死亡超過(guò)20萬(wàn)人。傷寒包括傷寒沙門(mén)氏菌(Salmonella typhi)引起的傷寒和甲、乙、丙型副傷寒沙門(mén)氏菌(Salmonella paratyphi A，，B and C)引起的副傷寒。近年來(lái)我國(guó)甲型副傷寒的發(fā)病率呈逐年上升趨勢(shì)，研究出安全、高效、經(jīng)濟(jì)的疫苗，對(duì)于預(yù)防甲型副傷寒爆發(fā)具有重大的經(jīng)濟(jì)社會(huì)效益。 甲型副傷寒沙門(mén)氏菌是無(wú)莢膜的革蘭氏陰性菌，其外膜蛋白在致病和刺激機(jī)體免疫應(yīng)答方面起著非常重要的作用。因此，在研究和開(kāi)發(fā)疫苗中，病原體的外膜蛋白成為熱點(diǎn)。 本論文對(duì)甲型副傷寒沙門(mén)氏菌外膜蛋白的克隆表達(dá)及其免疫保護(hù)性進(jìn)行了研究，目的在于探索應(yīng)用甲型副傷寒沙門(mén)氏菌外膜蛋白作為保護(hù)性抗原，獲得甲型副傷寒候選疫苗的可能性。 本課題選擇毒力較強(qiáng)的甲型副傷寒沙門(mén)氏菌50973做為實(shí)驗(yàn)菌株，以NmpC、MipA、OmpW、PagC、OmpX、PomP(GI：56412276)、OmpC、FadL、TolC、BtuB和Imp等11個(gè)外膜蛋白為目標(biāo)蛋白，分別將其基因的PCR擴(kuò)增片段克隆到pET-32a表達(dá)載體，篩選出陽(yáng)性克隆，用IPTG作為誘導(dǎo)劑，實(shí)現(xiàn)了這11種目標(biāo)外膜蛋白的過(guò)量表達(dá)。利用這些過(guò)表達(dá)目標(biāo)蛋白在鹽溶液中形成沉淀而在水中可溶的特性實(shí)現(xiàn)了目標(biāo)蛋白的有效分離。采用腹腔注射和體內(nèi)攻毒的方法對(duì)純化好的外膜蛋白的免疫保護(hù)性進(jìn)行了研究，結(jié)果表明，PagC、OmpX、NmpC、FadL、TolC和BtuB等6個(gè)外膜蛋白具有良好的免疫保護(hù)力，其保護(hù)率分別為100％、100％、95％、75％、75％和70％。
[Abstract]:Typhoid is an acute systemic infection that affects about 22 million people worldwide each year with typhoid fever (including paratyphoid fever). More than 200,000 people were killed. Typhoid includes typhoid fever caused by Salmonella typhimurium (Salmonella typhimurium). Paratyphoid fever caused by Salmonella paratyphi Agna B and (C). In recent years, the incidence of paratyphoid A in China has been increasing year by year. The study of a safe, efficient and economical vaccine has great economic and social benefits for the prevention of paratyphoid A (paratyphoid A) outbreak. Salmonella paratyphoid A is a gram-negative bacteria without capsule, and its outer membrane protein plays a very important role in pathogenic and stimulating immune response. Therefore, in the research and development of vaccines. The outer membrane protein of pathogens has become a hot spot. In this paper, the cloning and expression of the outer membrane protein of Salmonella paratyphi A and its immune protection were studied. The purpose of this study was to explore the application of the outer membrane protein of Salmonella paratyphoid A as a protective antigen. The possibility of obtaining a candidate vaccine for paratyphoid A (paratyphoid A). In this study, the virulent Salmonella paratyphoid A (50973) was selected as the experimental strain, and NmpCT-MipAmpWPagCor OmpX was used as the experimental strain. Pom Pi: 56412276OmpCy FadLn TolCtuB and Imp were the target proteins. The PCR fragments of the gene were cloned into the pET-32a expression vector, and the positive clones were screened out. IPTG was used as the inducer. The overexpression of the 11 target outer membrane proteins was realized. The effective separation of the target proteins was achieved by intraperitoneal injection and in vivo, using the characteristics of the over expressed proteins precipitating in salt solution and soluble in water. The immunological protection of purified outer membrane protein was studied by the method of virus attack. The results showed that the six outer membrane proteins, such as PagCon OmpX BtuB, Fad LT TolC and BtuB, had good immunological protection, and the protective rates were 100% and 100%, respectively. There are 75 percent and 70 percent.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R378

【引證文獻(xiàn)】

相關(guān)期刊論文 前2條

1 高翔;閆梅英;闞飆;;甲型副傷寒沙門(mén)菌疫苗靶標(biāo)蛋白篩選技術(shù)及應(yīng)用[J];疾病監(jiān)測(cè);2011年07期

2 陳金龍;王希輝;王婷婷;胡敬東;趙宏坤;許瑞利;;禽腸炎沙門(mén)菌外膜蛋白OMPX基因的克隆及其在乳酸乳球菌中的表達(dá)[J];中國(guó)獸醫(yī)學(xué)報(bào);2011年04期

本文編號(hào)：1413414