Expression Of SARS Coronavirus Structural Proteins And Its DNA Vaccine Construction

A novel coronavirus (SARS-coronavirus, SARS-CoV) was discovered the pathogen of the severe acute respiratory syndrome (SARS). Besides, the four main structural proteins play a mainly role for virus assemble and cell infection. It is important for SARS diagnosis reagent exploitation and Vaccine construction to express the four main structural proteins.To study the immunological speciality of structual proteins of SARS-CoV and analyze the feasibility and necessity of protein as the component of the SARS vaccine.The proteins of SARS-CoV was expressed in E.coli by pET-15b , pET-22b and pQE-2 and then purified by affinity resin as antigen . The four genes were cloned into the eukaryotic secretory expression plasmid pSecTagB and named recombinant plasmid pSecM, pSecS, pSecN, pSecE as DNA vaccine which will be used to immunize BALB/c mice to obtain the antiserum of M protein. The immune effect of the DNA vaccine was studied by the ELISA technique established with purified recombinant proteins. Both of the recombinant proteins can be expressed in soluble form. When compared with the SARS-CoV ELISA kit produced by Huada Co.Ltd, the prokaryotic reconbinant proteins could react specifically with the sera from immunized DNA mice and SARS correct diagnosis patients. The recombinant proteins could be used as a antigen to detect SARS and the SARS-CoV DNA vaccine could induce specifically antibody. Recombinant proteins could be considered as SARS-CoV vaccine especially as the vaccine component, which redounds to the development of a potential DNA vaccine.