Recombination and Expression of SARS Coronavirus Envelop Protein and Generation of Specific Monoclonal Antibody

To express small envelop(E) protein of SARS coronavirus and produce monoclonal antibody(mAb) to E protein. E protein gene was amplified by PCR. After being confirmed by DNA sequencing and linked with human immuno global protein gene Fc segment, the fused gene was sub-cloned into prokaryotic expression vector, pPRO EX Hta. EFc protein expressed in E.coli was recovered from SDS-PAGE gel and served as immunogen in the preparation of the mAb. Result of DNA sequencing confirmed that the amplified fragment was E protein gene. SDS-PAGE analysis showed that the expressed protein in E.coli was located at approximately Mr being 37×103 on the gel. This protein could be further confirmed by Western blot with serum of a SARS patient. The recombinant EFc protein was recovered from SDS-PAGE gel and immunized in BALB/c mice. Western blot analysis proved that two mAbs obtained could reacted specifically to the recombinant EFc protein. The recombinant EFc protein and mAbs lay the foundation for further in development of early diagnosis of SARS coronavirus infection.