Expression Of SARS Coronavirus Spike Protein In E.coli And Bombyx Mori

The part sequence (Base pairs from 22908 to 23542 in SARS-CoV genome sequence whose accession number is AY304495) of spike protein gene of SARS Coronavirus was cloned into the expression vector pET28a (+). SDS-PAGE and Western Blotting demonstrated that in E.coli BL21 recombinant S protein gene was expressed with the molecular weight about 27kDa of identity of its theoretical molecular weight. Also this part sequence of spike protein gene was inserted into baculovirus transfer vector pBacPAK-His, then cotransfected with liner silkworm baculovirus BmBacPAK-6 DNA in BmN Cells to construct the recombinant baculovirus BmBac-S. Plaque assay was taken to screen it. SDS-PAGE and Western Blotting analysis performed that the molecular weight of recombinant S protein was about 30kDa in BmN cell, and at 5 days post-infection its expression level could reach the maximum of about 1.2% of total hemolymph protein. The pure recombinant S protein expressed by baculovirus could be easily obtained through one-step metal chelate affinity chromatographer.