Construction and expression of SARS-CoV gene recombinant plasmids

Objective: To construct a recombinant eukaryotic expression plasmids pVAX1/SARS-CoV M, N, S, E based on their protein coding-gene, COS-7 cells were transfected by these recombinant plasmids, which provide scientific evidence for preparing SARS DNA vaccine. Methods: SARS-CoV M, N, S, E genes were amplified from Vero-E6 cells infected with SARS-CoV by RT-PCR. They were inserted into the eukaryotic expression vectors called pVAX1-M, pVAX1-N, pVAX1-S, pVAX1-E; COS-7 cells were transfected with recombinant plasmids; M, N, S, E protein were detected by Immunocytochemisty method and Western blot. Results: Recombinant plasmids were confirmed by restriction and DNA sequencing; M, N, S, E proteins were detected in COS-7 cells. Conclusion: All recombinant eukaryotic expression plasmids have been successfully constructed.