The Development Of Engineer Of Genetic Engineering Vaccine Of Procine Epidemic Diarrhea Virus

Porcine Epidemic diarrhea(PED),which is caused by Porcine Epidemic Diarrhea virus(PEDV)with a high fatality rate,is one of the most threatening swine diseases in the world.The outbreak of the disease around the world has caused significant losses to the global swine industry,but so far there is no effective and safe vaccine available.Thus,it is necessary to develop safe and efficient PEDV vaccine.Protein S is the main structural antigen protein of PEDV,which consists of S1 region and S2 region.S1 region,contains 3 neutralizing epitopes,is often an important target protein in the study of PEDV genetic engineering vaccine.In this study,recombinant baculoviruses contain different copies of S1 gene were constructed by using MultiBac expression system: AcMNPV-IM-p10S1,AcMNPV-IM-2p10S1,AcMNPV-IM-3p10S1,AcMNPV-IM-phS1,AcMNPV-IM-2phS1,AcMNPV-IM-3phS1,AcMNPV-IM-p10S1-phS1 and AcMNPV-IM-2p10S1-2phS1 to achieve efficient expression of S1 protein in insect cells.The S1 protein was used as a genetic engineering inactivated vaccine,and the immunological effect of the vaccine on the BALB/c mice model was preliminarily explored.Through the qualitative analysis of Western Blot and quantitative detection with his labeled ELISA kit,the results showed that after Sf9 cells were infected with different copies of recombinant baculoviruses,S1 were expressed efficiently in insect cells.1.When the number of S1 copies increased from 1 copy to 3 copies,the expression increased with the number of copies,3 copies of the expression reached the highest,4 copies of the decrease.2.The protein expression level drived by p10 promoter was significantly higher than that drived by p10 promoter polh promoter.S1 recombinant protein was prepared to immunize the dorsal subcutaneous immune mice.Indirect ELISA was carried out to detect the immune effect of immunological stimulation in mice.The results showed that the recombinant S1 protein could stimulate the immune response in mice.This research has created a new idea for the research and development of genetic engineering vaccines based on S-protein,which provides strong support for the clinical diagnosis and detection of swine epidemic diarrhea disease.