Expression of SARS-CoV Spike Protein Fragment of S144-643 and Its Immunogenicity

The gene fragment of S144-643 was subcloned into pET-28b and expressed the protein in E.coli.The protein existed mainly in bacterial inclusion bodies.The S144-643 was purified from bacterial inclusion bodies,refolded with the refolding buffer containing reduced glutathione and oxidized glutathione.SDS-PAGE demonstrated the protein with the molecular mass of about 55 ku.The immunogenicity of S144-643 was demonstrated by ELISA and Western blot assay,while the recombinant protein was specifically recognized by the sera from convalescent SARS patients.Furthermore,S144-643 was able to induce high titer of specific antibodies in immunized New Zealand rabbits.The S144-643 protein was expressed in E.coli and was immunologically active.