Expression And Identification Of E And M Protein Of Porcine Epidemic Diarrhea Virus

Porcine epidemic diarrhea(PED) is an acute, highly infectious intestinal disease,caused by porcine epidemic diarrhea virus(PEDV). The outbreak of PED in Asia is acute and the mortality rate is high, which it has had a great economic impact on the Asian pork industry. Vaccination of sows is a fundamental tool in a strategy to control and eradicate PED during epidemic or endemic outbreaks, but so far has not developed a particularly effective vaccines can control the occurrence of PED disease.Therefore, there was an urgent need for a new vaccine.PEDV is a large-enveloped RNA virus, and its genome is approximately 28 kb long. The study found that PEDV membrane glycoprotein(m) can elicit the production of protective antibodies with virus-neutralizing activity, and co- expression of membrane glycoprotein(m) and membrane protein(E) also can induce interferon production and enhance the antiviral ability. Therefore, M and E protein can be a candidate antigen for PEDV gene engineering vaccine.In this study, E and M amplified fragments were cloned into prokaryotic expression vector p ET-32 a, recombinant vectors was transformed into E.coli BL21 cells and induced by IPTG for expression. And E, M protein were purified and used to immunize BALB/c mice to prepared E, M polyclonal antibodies. PEDV E, M amplified fragments also cloned into pfastbac-dual vector, which expressed E, M protein. Using the polyclonal antibody to detect co-expression proteins. The results showed that E and M were expressed in Sf9 cells, and has good reactivity with preparation of polyclonal antibody.In this study, E, M protein was successfully expressed by the eukaryoticexpression vector—insect baculovirus expression system, which provided the basis for the further research of PEDV vaccine.