嗜水气单胞菌外膜蛋白基因DNA疫苗载体的构建及分析  

Construction and Characterization of DNA Vaccine for Major Outer Membrane Protein Gene of Aeromonas hydrophila

作　　者：李盼[1,2] 李素一[2] 吴唯维[2,3] 林天龙[2] 林晨韬[2] 陈叙[2] 

机构地区：[1]福建农林大学,福建福州350002 [2]福建省农业科学院生物技术研究所,福建福州350003 [3]福建师范大学,福建福州350007

出　　处：《福建农业学报》2013年第9期844-848,共5页Fujian Journal of Agricultural Sciences

基　　金：福建省农业科学院博士科研启动基金项目(2010BS-4);福建省农业科学院引进海外人才科研启动基金项目(HWRC2011-03);福建省自然科学基金项目(2011J05055);国家自然科学基金项目(31100658)

摘　　要：为构建嗜水气单胞菌的DNA疫苗载体,根据已发表的该菌外膜蛋白基因momp的核苷酸序列设计一对特异性引物,应用PCR技术,扩增到嗜水气单胞菌L316的主要外膜蛋白基因,并插入到真核表达载体pcDNA3上,构建成DNA疫苗,命名为pcDNA3-POMP。以纯化的原核表达蛋白GST-POMP免疫SD大鼠制备抗血清,用ELISA测定抗体效价达到1∶100 000以上;用pcDNA3-POMP质粒转染293细胞,转染48h后收集细胞,提取细胞的RNA进行RT-PCR,检测到外源基因的表达。至此,初步完成嗜水气单胞菌DNA疫苗表达载体的构建,为进一步疫苗免疫和效价的检测奠定基础。For constructing the DNA vaccine , a pair of specific PCR primers were designed according to the nucleotide sequence of Aeromonas hydrophila major outer membrane protein gene published by our lab previously . The PCR products were inserted into the eukaryotic expression vector pcDNA 3 to construct a DNA vaccine named pcDNA3-POMP . SD rats were immunized with purified prokaryotic-expressed protein GST-POMP to make antiserum ,whose antibody titer ,measured by ELISA ,was over 1 ∶ 100 000 .293 cells were transfected with pcDNA3-POMP ,48 hours after transfection ,the cells were collected for total RNA extraction and RT-PCR ,and the expression of exogenous momp were detected .Therefore ,we have constructed a plasmid of DNA vaccine for Aeromonas hydrophila ,vaccination and evaluation of this DNA vaccine will be studied in the future .

关 键 词：嗜水气单胞菌 DNA疫苗 外膜蛋白 

分 类 号：S941[农业科学—水产养殖]