Yeast Vaccine Design,Preparation And Detection Of African Swine Fever Virus

African swine fever is a severe infectious disease caused by African Swine Fever Virus（ASFV）.Pigs will die within 7–10 days after being infected with virulent strains.With the gradual deepening of virus research,the analysis of virus structure,the infection mode of virus has also been understood.It has been found that pigs infected with ASFV and survived have immunity to the virus,which makes people believe that vaccines will be an effective way to prevent African swine fever virus.In previous studies,for the design of African Swine Fever Virus vaccines,from attenuated vaccines to subunit vaccines to m RNA vaccines,the selection of viral proteins is also diverse.However,none of these vaccines have achieved satisfactory experimental results.The research of Bruno Elnaz and others confirmed that the infection of the virus is mainly non-specific macropinocytosis,so simply mediated antibody immunity will not have a good protective effect against African Swine Fever Virus attack.It is speculated that strong enough cellular immunity is required to have a protective effect.Previous studies in our laboratory have shown that gavage of whole yeast vaccine has delivered antigens to mouse intestinal epithelial cells,but subsequent flow cytometry tests found that whole yeast vaccine did not induce cellular immunity in mice.Studies in recent years have shown that after yeast strains are phagocytosed by macrophages,m RNA in yeast strains will be released into macrophages and translated and expressed in macrophages.In this study,the CoⅠ-P72s-αlectin/IRES-P72s plasmid was constructed according to Sun Young Sunwoo’s research and electroporated into the X33-PDI engineered yeast strain.Yeast genome PCR was performed to verify the construction of the strain and methanol induction was performed on the verified strain to confirm that the target gene could be expressed normally.Yeast strains were induced in YPM medium by adding methanol for 3 days and made into lyophilized powder.The lyophilized powder was divided to ensure that each tube contained 1.5×10~9 yeasts.BALB/c mice were gavaged every other week for one week for 3 months.The results of humoral immunity and cellular immunity of mice were detected.