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Virologica Sinica, 20 (1) : 1-4, 2005
Research Article
Cloning and Expression of SARS Coronavirus M gene
The full-length gene of membrane protein ( M )of SARS coronavirus (SARS-CoV) was cloned using RT-PCR from the Vero-E6 cells infected with SARS-CoV (GD322 strain) and inserted into the multi-cloning site of the pPICZαB vector. The recombinant plasmid was transtormed into P. pastoris strain X-33 by electroporation and selected by Zeocin. Mut phenotype determination was performed on the yeast transformants and then expression of Mut~+ colonies were induced by methanol. The expressed products were analyzed by SDS-PAGE and Western blotting. Secreted expression was performed by screening Mut~+ colonies in the yeast (transformants). The molecular mass of the recombinant M protein was approximately 65 kDa and 42 kDa and secreted into culture medium when induced with methanol. The expressed protein was able to (react) with SARS convalescence polyclonal antibody.
  Published online: 20 Jan 2005
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