【摘要】： 海豚鏈球菌是一種革蘭氏陽(yáng)性菌,是水產(chǎn)養(yǎng)殖業(yè)重要的病害之一,能夠感染多種水產(chǎn)魚類甚至人類。本研究運(yùn)用體內(nèi)誘導(dǎo)抗原技術(shù)從海豚鏈球菌中鑒別出一個(gè)分泌型抗原,Sia10。運(yùn)用CMV真核啟動(dòng)子,構(gòu)建DNA疫苗pSia10,用大菱鲆作為動(dòng)物模型研究其免疫保護(hù)性。實(shí)驗(yàn)表明在免疫后7D,14D,21D,28D,35D,42D,49D時(shí),Sia10在魚的肌肉,肝,脾,腎中均有表達(dá)。另外,免疫膠體金技術(shù)證明Sia10蛋白在免疫魚的肌肉中表達(dá)。免疫學(xué)分析表明免疫pSia10引起頭腎巨噬細(xì)胞產(chǎn)生高水平的氧化氮產(chǎn)物,酸性磷酸酶活性和化學(xué)發(fā)光反應(yīng)。熒光定量PCR表明免疫相關(guān)基因IgM, C3, Mx, NKEF, TLR3和TNF-α都有大于3倍的上調(diào)。致死率分析表明免疫Sia10 DNA疫苗的大菱鲆的死亡率顯著低于對(duì)照組,用不同劑量SF1攻毒檢測(cè)其相對(duì)保護(hù)效率可以達(dá)到73.9%和92.3%。這些結(jié)果表明,pSia10可作為一個(gè)有效的抵御SF1感染的候選疫苗。
[Abstract]:Streptococcus dolphin is a Gram-positive bacteria and one of the most important diseases in aquaculture. In this study, a secretory antigen, Sia10., was identified from Streptococcus dolphin by in vivo induced antigen technique. DNA vaccine pSia10, was constructed using CMV eukaryotic promoter as an animal model to study the immune protection of turbot. The results showed that Sia10 was expressed in muscle, liver, spleen and kidney of fish after immunization. In addition, immuno-colloidal gold technique demonstrated that Sia10 protein was expressed in the muscle of immunized fish. Immunological analysis showed that immune pSia10 induced high level of nitrogen oxide production, acid phosphatase activity and chemiluminescence reaction. Fluorescence quantitative PCR showed that the immune-related genes IgM, C _ 3, Mx, NKEF, TLR3 and TNF- 偽 were up-regulated by more than three times. The mortality analysis showed that the mortality of turbot immunized with Sia10 DNA vaccine was significantly lower than that of the control group. The relative protective efficiency of different doses of SF1 was 73.9% and 92.3% respectively. These results suggest that pSia10 can be an effective candidate vaccine against SF1 infection.
【學(xué)位授予單位】：中國(guó)科學(xué)院研究生院（海洋研究所）
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R392

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