Construction Of NC/TGAMA1 Gene Of Cross Antigen And Recombinant Adenovirus Vector Vaccine And Animals Immunity

Both Neospora caninum and Toxoplasma gondii were have closed relationship,all belong to the intracellular parasitic protozoa,and so both were very similar in morphology,molecular biological characteristics and clinical symptoms and disease,and infected can lead to miscarriage Yunxu,stillbirth,mummified and other symptoms.AMA1 apical membrane antigen labeled in local micro-line body,expressed in Neospora caninum and Toxoplasm gondii tachyzoites gentle son of colonization with parasites invaded host cells and copied functions were closely related.Study confirmed,AMA1 protein can be effectively controlled Neospora caninum and Toxoplasma gondii parasited invasion,Neospora caninum and Toxoplasma gondii were apotential candidate vaccine antigens.Currently,Neospora caninum and Toxoplasma gondii has no effective prevention and control of drugs and vaccines.This test based on Neospora caninum and Toxoplasma gondii AMA1 gene sequence of the open reading frame,designed Neospora caninum and Toxoplasmagondii antigen AMA1 gene crossed universal primers.Constructed pMD 18-T-Nc/TgAMA1 recombinant plasmid cloning,recombinant adenovirus shuttle plasmid ADV4-Nc/TgAMA1,ADV4-Nc/Tg AMA1 and backbone plasmid linearized pacAd5 293T cells were co-transfected packaged Ad5-Nc/TgAMA1 adenovirus,amplified and purified were prepared Ad5-Nc/TgAMAl recombinant adenoviral vector vaccine.Inoculated BALB/c mice,and set pVAX1-Nc/TgAMAl plasmid group and PBS group,ELISA assay vaccinated mouse serum IgG antibody levels and IFN-γ,IL-4 cytokine levels.Two weeks after the third immunization,each group of BALB/c mice were divided into two groups,they were given intraperitoneal attack Neospora caninum and Toxoplasma gondii,clinical symptoms and survival statistics mice.The results displayed,obtained Ad5-Nc/Tg AMA1 recombinant adenovirus titer of 109 PFU/mL,after inoculation BALB/c mice,IgG antibody levels,Ad5-Nc/TgAMAl vaccine group was significantly higher than pVAXl-Nc/TgAMAl plasmid PBS group and the control group(P<0.01);IFN-Y and IL-4 cytokines,Ad5-Nc/TgAMA1 vaccine group were significantly higher than pVAXl-Nc/TgAMAl plasmid group and PBS control group(P<0.01).Attack by insects tests showed that vaccinated Ad5-Nc/TgAMA1 vaccine BABL/c mice Neospora caninum and Toxoplasma gondii attacked were 80%and 60%survival rate,pVAX1-Nc/TgAMAl plasmid group were 50%and 20%survival rate,while the PBS control group were 20%and 0%survival.Description The experiment produced Ad5-Nc/TgAM1 vaccine against Toxoplasma gondii and Neospora caninum attacked with good immune protection,achieved while preventing Neospora caninum and Toxoplasma gondii infected purposes.