Article Figures & Data
Figures
- FIG. 1.
(A) Expression of the N protein of SARS-CoV in E. coli. Lanes 1 to 12, SDS-PAGE of bacterial cell lysates harvested at 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 18 h, respectively, after IPTG induction; lane 13, soluble proteins in the supernatant of cell lysates; lane 14, SDS-PAGE of 5 μg of the purified N protein. (B) Western blot of soluble proteins in the supernatant (B1) and of the purified protein (B2) using monoclonal antibody to the Xpress epitope tag fused with the N protein. A molecular marker (M) is indicated. Arrows, 50 kDa.
- FIG. 2.
Western blot analyses of antigenic cross-reactivity of SARS-CoV N protein with polyclonal antisera of known animal coronaviruses. SARS-CoV N protein (1 μg/lane) was separated by SDS-PAGE. Each antiserum was diluted 1:100 in blocking solution. A convalescent-phase SARS human patient serum (anti-SARS-CoV) and anti-Xpress antibody were used as positive controls. The polyclonal antisera used in the Western blot analysis were from antigenic group I (FIPV, porcine TGEV, and CCoV), group II (porcine HEV and BCoV), and group III (TCoV and avian IBV) animal coronaviruses. Polyclonal antisera to IBV, HEV, BcoV (calf serum), and TGEV (pig serum) were purchased from National Veterinary Service Laboratories, Ames, Iowa. Polyclonal cat antisera to TGEV and CCoV and cat ascitic fluid against FIPV were purchased from VMRD, Inc., Pullman, Wash. The arrow shows the expected size (about 50 kDa) of the SARS-CoV N protein.
