Citation: YAN Hui-jun, ZHAO Wei, FANG Dan-yun, ZHOU Jing-jiao, LONG Bei-guo, ZHANG Wen-bing, GUO Hui-yu, JIANG Li-fang. Cloning and Expression of SARS Coronavirus M gene .VIROLOGICA SINICA, 2005, 20(1) : 1-4.

Cloning and Expression of SARS Coronavirus M gene

  • Available online: 20 January 2005
  • 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.

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    Cloning and Expression of SARS Coronavirus M gene

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    Abstract: 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.

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