【摘要】：絲蟲病是一種嚴重危害人類健康的傳染病,長期使用化學藥物,使絲蟲和蚊媒抗藥性增強。研究和發(fā)展價格低廉、安全高效的絲蟲疫苗是未來絲蟲病防治工作中極有潛力的重要課題。近年來發(fā)展起來的核酸疫苗以其制備簡便、實用高效等優(yōu)點而備受重視,具有廣闊的應(yīng)用前景�，F(xiàn)有的資料表明3-磷酸甘油醛脫氫酶(glyceraldehyde-3-phosphate dehydrogenase, GAPDH)在寄生蠕蟲和原蟲中,可誘導(dǎo)實驗動物產(chǎn)生保護性免疫應(yīng)答,是一種重要的疫苗候選抗原。本研究成功構(gòu)建了周期型馬來絲蟲核酸疫苗pcDNA3.1/Bm-gapdh。為了提高免疫效應(yīng),用免疫刺激序列CpG作為佐劑共同免疫BALB/c小鼠,用RT-PCR、MTT、ELISA檢測GAPDH的免疫效應(yīng)。結(jié)果表明,核酸疫苗pcDNA3.1/Bm-gapdh具有抗絲蟲病候選DNA疫苗分子的潛力,增加佐劑后免疫效應(yīng)有所增強。為了下一步的疫苗設(shè)計作前期準備,本實驗還構(gòu)建了周期型馬來絲蟲半胱氨酸蛋白酶抑制劑(cysteine protease inhibitor,CPI)的重組質(zhì)粒pGEM-T/Bm-cpi,并送基因公司測序。再根據(jù)測序結(jié)果,推測其氨基酸序列,然后利用相關(guān)的軟件預(yù)測它的B細胞表位。
[Abstract]:Filariasis is an infectious disease that seriously endangers human health. Long-term use of chemical drugs enhances resistance to filariasis and mosquito vectors. The research and development of low cost, safe and efficient filariasis vaccine is an important topic with great potential in filariasis control in the future. The nucleic acid vaccine which has been developed in recent years has attracted much attention because of its advantages of simple preparation, practical efficiency and so on, and has a broad application prospect. The available data show that glyceraldehyde-3-phosphate dehydrogenase, GAPDH) can induce protective immune response of experimental animals in parasitic worms and protozoa and is an important vaccine candidate antigen. In this study, we successfully constructed the pcDNA3.1/Bm-gapdh. of periodic filariasis nucleic acid vaccine. In order to improve the immune effect, BALB/c mice were immunized with CpG as adjuvant, and the immune effect of GAPDH was detected by RT-PCR,MTT,ELISA. The results showed that the nucleic acid vaccine pcDNA3.1/Bm-gapdh had the potential to be a candidate DNA vaccine against filariasis, and the immune effect was enhanced after increasing the adjuvant. In order to prepare the vaccine for the next step, the recombinant plasmid pGEM-T/Bm-cpi, of cysteine protease inhibitor (cysteine protease inhibitor,CPI (Cysteine protease inhibitor) of filariasis malayi was constructed and sequenced. The amino acid sequence was deduced according to the results of sequencing, and then the B cell epitopes were predicted by the relevant software.
【學位授予單位】：南通大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R392

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