本文選題：衣殼蛋白 + VP1��； 參考：《上海交通大學(xué)》2011年碩士論文

【摘要】：人腸道病毒71型(EV71)屬于小RNA病毒科、腸道病毒屬的成員,是手足口病的主要病原體。手足口病和中樞神經(jīng)感染是EV71病毒感染引起的兩大常見臨床癥狀。目前針對(duì)EV71病毒感染的治療還沒有特異和高效的藥物,也沒有預(yù)防性疫苗上市。滅活疫苗的研究還處于Ⅰ期臨床試驗(yàn)階段;基因工程重組蛋白疫苗尚處于動(dòng)物實(shí)驗(yàn)階段,需要尋找好的有效抗原蛋白。因此,本論文在原核表達(dá)系統(tǒng)E.coli中高效表達(dá)了EV71四種衣殼蛋白:VP1、VP2、VP3、VP4,表達(dá)產(chǎn)物均以包涵體形式存在;包涵體經(jīng)過純化、復(fù)性后的SDS-PAGE電泳純度平均達(dá)到90%以上。動(dòng)物實(shí)驗(yàn)結(jié)果顯示VP1、VP4蛋白具有良好的免疫原性,產(chǎn)生高抗體滴度;VP2、VP3無明顯的免疫原性,沒有檢測到抗體滴度;僅VP1免疫血清具有中和活性。通過本論文實(shí)驗(yàn),加深了對(duì)EV71四種衣殼蛋白的認(rèn)識(shí);證實(shí)了VP4因?yàn)闆]有產(chǎn)生中和抗體,不具有成為基因工程重組蛋白疫苗候選抗原的潛力。同時(shí)證實(shí)了原核表達(dá)的VP2、VP3抗原蛋白經(jīng)過去變性劑初步復(fù)性后,與EV71病毒顆粒表面的VP2、VP3蛋白在空間構(gòu)象上有差異,需要進(jìn)一步深入研究;證實(shí)了VP1具有成為基因工程重組蛋白疫苗候選抗原的特性。這為今后EV71疫苗候選抗原的篩選提供了重要依據(jù)。
[Abstract]:Human enterovirus 71 (EV71), a member of the genus RNA, is a major pathogen of hand, foot and mouth disease. Hand, foot and mouth disease and central nervous system infection are two common clinical symptoms caused by EV71 virus infection. Currently, there are no specific and effective drugs for the treatment of EV71 infection, and no prophylactic vaccine is available. The research of inactivated vaccine is still in the stage of phase I clinical trial, and the recombinant protein vaccine is still in animal experiment stage, so it is necessary to search for a good effective antigen protein. Therefore, in the prokaryotic expression system E.coli, the four capsid proteins of EV71, 1 / VP1 / VP2 / VP3 / VP4, were highly expressed in the prokaryotic expression system, and all the expressed products were in the form of inclusion bodies, and the purity of SDS-PAGE after renaturation was over 90% on average. The results of animal experiments showed that VP1mVP4 protein had good immunogenicity, and the antibody titer of VP2VP3 was not detected, but that of VP1 immunized serum had neutralizing activity. Through the experiments in this paper, four kinds of capsid proteins of EV71 were further recognized, and it was confirmed that VP4 did not have the potential to be a candidate antigen for recombinant protein vaccine because it did not produce neutralizing antibody. At the same time, it was confirmed that the VP2VP3 protein expressed in prokaryotic cells was different from the VP2VP3 protein on the surface of EV71 virus after denaturalization, and needed further study. It is confirmed that VP1 is a candidate antigen for recombinant protein vaccine. This provides an important basis for the screening of EV71 vaccine candidate antigens in the future.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R373

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本文編號(hào)：1860348