海豚链球菌DNA疫苗的构建及其免疫效应分析

发布时间：2019-02-17 18:44

【摘要】： 海豚链球菌是一种革兰氏阳性菌,是水产养殖业重要的病害之一,能够感染多种水产鱼类甚至人类。本研究运用体内诱导抗原技术从海豚链球菌中鉴别出一个分泌型抗原,Sia10。运用CMV真核启动子,构建DNA疫苗pSia10,用大菱鲆作为动物模型研究其免疫保护性。实验表明在免疫后7D,14D,21D,28D,35D,42D,49D时,Sia10在鱼的肌肉,肝,脾,肾中均有表达。另外,免疫胶体金技术证明Sia10蛋白在免疫鱼的肌肉中表达。免疫学分析表明免疫pSia10引起头肾巨噬细胞产生高水平的氧化氮产物,酸性磷酸酶活性和化学发光反应。荧光定量PCR表明免疫相关基因IgM, C3, Mx, NKEF, TLR3和TNF-α都有大于3倍的上调。致死率分析表明免疫Sia10 DNA疫苗的大菱鲆的死亡率显著低于对照组,用不同剂量SF1攻毒检测其相对保护效率可以达到73.9%和92.3%。这些结果表明,pSia10可作为一个有效的抵御SF1感染的候选疫苗。
[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- 伪 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.
【学位授予单位】：中国科学院研究生院（海洋研究所）
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392

【参考文献】