Analysis of severe acute respiratory syndrome coronavirus structural proteins in virus‐like particle assembly |
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| Authors: | Mina Nakauchi Hiroaki Kariwa Yasuhiro Kon Kentaro Yoshii Akihiko Maeda Ikuo Takashima |
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| Institution: | 1. Laboratory of Public Health, Department of Environmental Veterinary Sciences;2. Laboratory of Anatomy, Department of Biomedical Sciences;3. Laboratory of Prion Diseases, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060‐0818, Japan |
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| Abstract: | SARS‐CoV has four major structural proteins: the N, S, M, and E proteins. To investigate the mechanism of SARS‐CoV assembly, we cloned the genes encoding these four proteins into the eukaryotic expression vector pCAGGS and transfected them into 293T cells. When all four expression vectors were co‐transfected VLP formed, as confirmed using electron microscopy. Using a rabbit polyclonal antibody specific to the N protein, N‐protein‐containing particles similar in size to the VLP were also observed by immunoelectron microscopy, indicating that the VLP contained the N protein. Co‐immunoprecipitation analyses demonstrated an interaction between the N and M proteins, suggesting that N protein binds directly to M protein to be incorporated into VLP. |
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| Keywords: | particle formation SARS‐CoV VLP |
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