Fig. 1.

Characteristics of SARS-CoV spike protein-expressing recombinant vaccinia virus (RVV-S) derived from LC16m8. (A) The full length SARS-CoV spike protein gene was inserted into the HA gene region of the LC16m8 genome. The ATI/p7.5 hybrid promoter regulates expression of spike protein. (B and C) RK13 cells were infected with RVV-S or LC16m8 at moi 10. At 24 h after infection, cells were harvested and analyzed. Two kinds of anti-SARS-CoV spike protein polyclonal antibodies, which recognize different epitopes, namely amino acid residues 559–570 (B) and 1236–1248 (C) of spike protein, were used as the primary antibodies. The molecular masses of marker proteins in kDa are shown on the left and the position of the spike protein is indicated by an arrowhead on the right. (D) Indirect immunofluorescence staining of spike protein. Expression of spike protein was visualized by staining with anti-SARS-CoV spike polyclonal antibodies, followed by Alexa 488-conjugated anti-rabbit IgG (green). Nuclei were stained with DAPI (red).