Expression of SARS Coronavirus Membrane Protein and its DNA Vaccine Construction

To study the immunological speciality of M protein of SARS-CoV and analyze the feasibility and necessity of M protein as the component of the SARS vaccine. M protein of SARS-CoV was expressed in E.coli by pET-15b and pET-22b and purified by affinity resin as antigen. M gene was cloned into the eukaryotic secretory expression plasmid pSecTagB and named recombinant plasmid pSecM as DNA vaccine which will be used to immunize BALB/c mice to obtain the antiserum of M protein. The immune effect of the M-DNA vaccine was studied by the ELISA technique established with purified M protein. Both of the recombinant M proteins can be expressed in soluble form. When compared with the SARS-CoV ELISA kit produced by Huada Co.Ltd, the prokaryotic reconbinant M proteins could react specifically with the sera from immunized M-DNA mice and SARS correct diagnosis patients. The recombinant M protein could be used as an antigen to detect SARS, and the SARS-CoV M gene vaccine could induce specifically antibody. M protein could be