金黄色葡萄球菌FnbA配体结合区基因核酸疫苗的构建与免疫试验

Study on Construction and Immune Experiment of DNA Vaccine Expressing p Fnb A Gene of Staphylococcus aureus of cow mastitis

扩增了奶牛乳腺炎金黄色葡萄球菌纤粘连结合蛋白A(Fnb A)配体结合区基因,并将其克隆至真核表达载体p VAX1启动子下游,构建成真核表达质粒,通过体外细胞转染试验,运用IFA方法进行抗原性初步确认,所构建的重组DNA疫苗质粒能在真核细胞中表达外源基因并被金黄色葡萄球菌抗体特异性识别。为进一步评价侯选疫苗的免疫原性,进行了BALB/c小鼠免疫试验,分别检测免疫后的ELISA抗体水平、Th1/Th2类细胞因子水平以及T淋巴细胞增殖试验。结果表明,构建的核酸疫苗p VAX1-p Fnb A免疫小鼠后,ELISA抗体水平提高,Th1/Th2类细胞因子含量提升,T细胞增殖能力增强。

In this study,Fnb A ligand attachment region D1-D3 gene（p Fnb A）of the Staphylococcus aureus was amplified and cloned into p VAX1 eukaryotic expressing vector. The recombinant DNA vaccine was transcribed and expressed in eukaryotic cells effectively through IFA identification.To further evaluate its immunogenicity,the DNA vaccine was used to immunize BALB/c mice.The ELISA antibody,number of Th1/Th2 cytokines and T cell multiplication were tested after immunization.The result shows that the ELISA antibody and the number of Th1/Th2 cytokine increased,and T cell multiplication was strengthened.