本文關(guān)鍵詞：乙型腦炎SA14-14-2株E、NS1蛋白的原核表達(dá)及其免疫原性研究　出處：《中國(guó)藥品生物制品檢定所》2006年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 乙型腦炎病毒 E、Ns1 ELISA 基因表達(dá) NS1蛋白保護(hù)效果

【摘要】：流行性乙型腦炎病毒(Japanese encephalitis virus，JEV)簡(jiǎn)稱乙腦病毒，又稱日本腦炎病毒，是一種嚴(yán)重危害人畜(尤其是兒童)健康的蟲媒病毒。目前沒有治療乙腦的有效藥物。接種乙腦疫苗是控制乙腦的最重要措施。我國(guó)是乙型腦炎發(fā)病人數(shù)最多的國(guó)家。但近十幾年來，乙腦的發(fā)病率明顯下降，其中原因主要?dú)w功于疫苗的接種。目前國(guó)際普遍接種的疫苗是鼠腦來源的滅活疫苗。如日本、韓國(guó)、新加坡、印度以及美國(guó)等使用鼠腦來源的Nakayama株和／或Beijing-1株的滅活疫苗。由于滅活疫苗存在一些缺點(diǎn)，即(1)需要多次接種才能產(chǎn)生良好的免疫效果，(2)不良反應(yīng)漸多，如局部反應(yīng)率達(dá)20％左右，全身反應(yīng)率達(dá)10％，(3)價(jià)格昂貴，生產(chǎn)工藝復(fù)雜等大大影響其進(jìn)一步推廣。因此需要研究生產(chǎn)更有效、安全、方便的疫苗。目前研究的新疫苗有兩類，第一為減毒活疫苗，由SA14衍生出的SA14-14-2已經(jīng)被多方面證明毒力低、安全性好。目前我國(guó)廣泛使用的疫苗有SA14-14-2株原代地鼠腎細(xì)胞減毒活疫苗和北京P3株地鼠腎細(xì)胞滅活疫苗。SA14-14-2株原代地鼠腎細(xì)胞減毒活疫苗在國(guó)內(nèi)已經(jīng)成功接種逾2億人，經(jīng)證實(shí)安全、有效、便宜。第二類為基因重組疫苗。IC47為無毒重組的JEV，但由于沒有解決穩(wěn)定劑型、清除毒力逆轉(zhuǎn)的問題，在作為正規(guī)疫苗應(yīng)用之前，有很長(zhǎng)的一段距離。因此，，減毒活疫苗SA14-14-2病毒株引起國(guó)內(nèi)國(guó)際疫苗界人士的高度重視。 本課題選用JEV SA14-14-2第一個(gè)被翻譯的非結(jié)構(gòu)基因ns1進(jìn)行研究，采用PCR方法擴(kuò)增NS1蛋白基因和E蛋白基因片段，將各基因片斷克隆到原核表達(dá)載體pET系統(tǒng)上，并使其在大腸桿菌中誘導(dǎo)表達(dá)，篩選出表達(dá)菌株pET-32Et和pET—30NS1，通過親和層析對(duì)兩種蛋白進(jìn)行純化，得到純度在96％以上的目的蛋白。
[Abstract]:Japanese encephalitis virus (je) is also known as Japanese encephalitis virus (JEV). Is a serious threat to people and animals (especially children). There is no effective drug to treat Japanese encephalitis. Vaccination with je vaccine is the most important measure to control Japanese encephalitis. China is the country with the highest incidence of encephalitis B, but in recent ten years. The incidence of encephalitis B has declined significantly, mainly due to vaccination. Currently, inactivated vaccines from the brain are widely used in the world, such as Japan, South Korea, and Singapore. Inactivated vaccines using brain-derived Nakayama and / or Beijing-1 strains in India and the United States. That is to say, many times of inoculation is needed to produce good immune effect. (such as local reaction rate of 20% or so, systemic reaction rate of 10 ~ 3%) is more and more expensive. The complexity of production technology has greatly affected its further promotion. Therefore, it is necessary to study and produce more effective, safe and convenient vaccines. There are two kinds of new vaccines, the first is live attenuated vaccine. SA14-14-2 derived from SA14 has been proved to be low virulence in many ways. At present, the widely used vaccines in China include SA14-14-2 strain primary hamster kidney cell attenuated vaccine and Beijing P3 strain hamster kidney cell inactivated vaccine .SA14-14-2 primary hamster kidney vaccine. Live attenuated cell vaccines have been successfully vaccinated against more than 200 million people in China. The second type is the gene recombinant vaccine. IC47 is the non-toxic recombinant JEV, but the problem of virulence reversal has not been solved because the stabilizer type has not been solved. Before being used as a regular vaccine, there is a long distance. Therefore, the attenuated live vaccine SA14-14-2 strain has attracted great attention from domestic and international vaccine researchers. In this study, the first translated non-structural gene ns1 of JEV SA14-14-2 was used to amplify NS1 gene and E protein gene by PCR. The gene fragments were cloned into the prokaryotic expression vector pET system and induced in Escherichia coli. The expression strains pET-32Et and pET-30NS1 were screened out. The two proteins were purified by affinity chromatography and the purity of the two proteins was over 96%.
【學(xué)位授予單位】：中國(guó)藥品生物制品檢定所
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 朱秀高;;國(guó)內(nèi)豬傳染性乙型腦炎的檢測(cè)及預(yù)防[J];養(yǎng)豬;2012年02期

相關(guān)碩士學(xué)位論文 前1條

1 戚龍霞;乙型腦炎病毒E蛋白在桿狀病毒表面的展示及其免疫原性研究[D];西北農(nóng)林科技大學(xué);2010年

本文編號(hào)：1376847