普通变形杆菌TWN3株DNA疫苗的初步研究  

DNA Vaccine of Bacterium Proteus vulgaris TWN3 Strain

作　　者：王娟[1,2] 辜文博[1] 冯军[1] 顾继锐[1,2] 吴江[2] 徐恒[1,2] 

机构地区：[1]四川大学生命科学学院,生物资源与生态环境教育部重点实验室,四川成都610064 [2]通威股份有限公司,四川成都610081

出　　处：《水产科学》2009年第4期205-208,共4页Fisheries Science

基　　金：四川省科技攻关项目(05NG002-002-4);通威股份科技发展项目

摘　　要：普通变形杆菌TWN3分离自死亡大口鲇,菌的DNA文库已构建成功,通过免疫印迹法从文库中筛选到TWN3抗原片段PV4,将其克隆到真核表达载体pcDNA3上,构建DNA疫苗pcDNA3-PV4。肌肉注射免疫小鼠进行真核表达检测。免疫组织化学分析DNA疫苗在接种局部肌肉中抗原的表达,ELISA检测小鼠血清中的IgG及IFN-γ含量。免疫pcDNA3-PV4疫苗的小鼠肌肉细胞中检测到PV4相关抗原蛋白的表达,外周血中的IgG及IFN-γ显著高于免疫pcDNA3组。对小鼠进行免疫保护试验,免疫DNA疫苗的小鼠对TWN3菌有免疫力。初步显示构建的DNA疫苗有一定效果。试验结果为鱼类水体试验提供了参考依据,同时随着TWN3株菌DNA疫苗研究的深入,将对该疫苗最终大规模用于鱼类提供理论基础。Bacterium Proteus vulgaris TWN3 strain was isolated from tissue of dead southern sheatfish Silurus meridionalis in which the DNA library was constructed successfully. PV4 coding the antigen was obtained from the DNA library by immunoscreening, then cloned into the vaccine vector pcDNA3 ,leading to establishment of DNA vaccine. Kunming mice were immunized through intramuscular inoculation. A immunohistobiochemical method was used to study the expression of specific antigen protein in the muscles. To detect IgG level and IFN-γ in sera,ELISA method was used. The results showed that specific antigen protein was expressed in the mice muscle immunized with pcDNA3-PV4, also IgG and IFN-γ level was largely different from the mice immunized with pcDNAa. Immunoproteetion experiment of mice indicated that the mice immuned with DNA vaccine were immuned against TWN3. The results provided scientific theory for the immunoprotection experiment of fish. As the experiment going on, it will be helpful for the DNA vaccine used largely in fish.

关 键 词：普通变形杆菌TWN3株 DNA文库 PV4 DNA疫苗 

分 类 号：Q78[生物学—分子生物学]