發(fā)布時(shí)間：2018-03-05 10:18

  本文選題：腦膜炎奈瑟氏菌表面蛋白A　切入點(diǎn)：克隆和表達(dá)　出處：《中國(guó)醫(yī)科大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】：腦膜炎奈瑟氏菌表面蛋白A基因的克隆和表達(dá) 前言 腦膜炎奈瑟氏菌是引起人流行性腦膜腦炎(以下簡(jiǎn)稱流腦)的致病菌。在非流行期,發(fā)病率在1-3/10萬(wàn)左右,而在流行期,發(fā)病可高達(dá)500/10萬(wàn),對(duì)6個(gè)月到2歲的嬰幼兒及青少年危害極大,病死率高達(dá)10%以上,近年來(lái),我國(guó)流腦發(fā)病率有明顯上升趨勢(shì),暴發(fā)流行時(shí)有發(fā)生,對(duì)我國(guó)人民的生命健康構(gòu)成極大威脅。 腦膜炎奈瑟氏菌根據(jù)菌體表面的莢膜多糖分為12個(gè)血清群。全世界大約90%的病人是由A群、B群和C群引起的。以A群、C群莢膜多糖為基礎(chǔ)的疫苗被開發(fā)出來(lái)并被證明對(duì)控制流腦的暴發(fā)和流行有效。但是A群、C群疫苗對(duì)幼兒缺乏免疫原性,而且它們提供的保護(hù)期相對(duì)短,一般為1-3年。B群莢膜多糖由于無(wú)法誘導(dǎo)人體產(chǎn)生免疫反應(yīng),所以B群流腦疫苗一直也沒有被開發(fā)出來(lái)。基于這些原因,開發(fā)非莢膜多糖的表面蛋白抗原疫苗以刺激機(jī)體產(chǎn)生保護(hù)性、持久性抗體就成為另一種選擇。 但是每個(gè)血清群的腦膜炎奈瑟氏菌根據(jù)菌體主要表面蛋白(OMP)又可分為多個(gè)血清型、血清亞型,如B群有20多個(gè)血清型,每個(gè)血清型又有至少10多個(gè)血清亞型,依此類推12個(gè)血清群的腦膜炎奈瑟氏菌有上千種表面蛋白。尋找所有血清群共有的表面蛋白抗原成為幾十年來(lái)全世界腦膜炎奈瑟氏菌研究者奮斗的目標(biāo),1997年Martin等發(fā)現(xiàn)了一種極為保守的表面蛋白,存在于所有的腦膜炎奈瑟氏菌菌體表面,命名為腦膜炎奈瑟氏菌表面蛋白A(NspA),含174個(gè)氨基酸,由525個(gè)核苷酸序列編碼組成。盡管Ns-pA的功能至今還不清楚,但給小鼠注射純化的或重組的NspA后都能誘導(dǎo)產(chǎn)生保護(hù)性抗體,抵抗腦膜炎奈瑟氏菌的攻擊。 綜上所述,NspA對(duì)流腦的診斷和預(yù)防策略的制定具有極大的應(yīng)用價(jià)值。為此,本研究對(duì)腦膜炎奈瑟氏菌表面蛋白A基因進(jìn)行了克隆和表達(dá),并采用ELISA對(duì)純化的NspA融合蛋白進(jìn)行了活性檢測(cè),同時(shí)驗(yàn)證了人體
[Abstract]:Cloning and expression of surface protein A gene of Neisseria meningitidis. Foreword. Neisseria meningitidis is the cause of human epidemic meningoencephalitis (hereinafter referred to as meningitis). It is harmful to infants and adolescents aged from 6 months to 2 years old, and the mortality is more than 10%. In recent years, the incidence of meningitis in China has an obvious upward trend, outbreaks and epidemics occur from time to time, which poses a great threat to the life and health of the people in our country. Neisseria meningitidis is divided into 12 serogroups based on capsular polysaccharides on the surface of the bacteria. About 90% of the world's patients are caused by groups A, B and C. Vaccine based on group A and group C capsule polysaccharides has been developed and developed. It has been proved to be effective in controlling the outbreak and epidemic of meningitis. But the group A and C vaccine is lack of immunogenicity in young children. And they offer relatively short protection periods, typically 1-3 years. Group B capsule polysaccharides have not been developed because they can't induce immune responses in humans. For these reasons, The development of a surface protein antigen vaccine for non-perfringent polysaccharides to stimulate protection in the body makes persistent antibodies an alternative. But each serotype of Neisseria meningitidis can be divided into several serotypes according to its main surface protein, serotypes, such as more than 20 serotypes in group B, and at least more than 10 serotypes in each serotype. And so on 12 serogroups of Neisseria meningitidis have thousands of surface proteins. Looking for surface protein antigens shared by all serums has been the goal of researchers around the world for decades, Martin et al., 1997. Found a very conservative surface protein, It exists on the surface of all Neisseria meningitidis. It is named as the surface protein of Neisseria meningitidis, AnspAN, which contains 174 amino acids and is composed of 525 nucleotide sequences, although the function of Ns-pA is still unknown. But injected with purified or recombinant NspA could induce the production of protective antibodies against Neisseria meningitidis. To sum up, NspA has great application value in the diagnosis and prevention of meningitis. Therefore, the surface protein A gene of Neisseria meningitidis was cloned and expressed in this study. The activity of purified NspA fusion protein was detected by ELISA, and the human body was verified.
【學(xué)位授予單位】：中國(guó)醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R378

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 鄭鑫;腦膜炎奈瑟氏菌表面蛋白NspA基因的克隆與原核表達(dá)[D];長(zhǎng)春理工大學(xué);2011年

，

本文編號(hào)：1569825