Table 2. Efficiency of virus generation in Vero cells
Experimental samples
Components/results 1 2 3 4 5 6 7 8 9 10 11 12 13 14
vRNA synthesis
8-unit plasmid + + +
6-unit plasmid + + + +
2-unit plasmid + + + +
8 × 1-Unit plasmid + + +
Protein synthesis
pCAWS-PB2 + + + +
pCAWS-PB1 + + + +
pCAWS-PA + + + +
pCAWS-NP + + + + + + + +
pC-PolII-WSN-PB2-PB1-PA + + + +
Total no. of plasmids 1 5 3 1 1 2 6 4 12 10 4 2 8 0
Exp. 1 (48 h p.t.), TCID50/ml <50 1.7 × 104 3.7 × 104 0 0 0 <10 3.2 × 104 <10 <50 0 0 0 0
Exp. 2 (48 h p.t.), TCID50/ml 0 <50 3.7 × 104 0 0 0 6.2 × 102 4.4 × 104 <10 1.5 × 103 0 0 0 0
Exp. 3 (48 h p.t.), TCID50/ml 0 3 × 102 5.1 × 104 0 0 0 1.6 × 102 2 × 104 2 × 103 2.5 × 103 0 0 0 0
Exp. 1 (72 h p.t.), TCID50/ml 3.2 × 104 6.3 × 105 2.5 × 106 0 0 0 5.3 × 103 3.9 × 105 2.5 × 103 6.3 × 104 0 0 0 0
Exp. 2 (72 h p.t.), TCID50/ml <10 3.1 × 103 1.6 × 106 0 0 0 7.6 × 104 6.3 × 105 6.3 × 103 2.1 × 105 0 0 0 0
Exp. 3 (72 h p.t.), TCID50/ml 50 3.2 × 104 2 × 106 0 0 50 2 × 104 5.1 × 105 1.6 × 105 2.5 × 105 0 0 0 0
  • Vero cells were transfected with the indicated plasmids (+). At 48 h or 72 h after transfection, virus titers in the supernatant were determined by plaque assays in MDCK cells. Shown are the results of three independent experiments. Eight-unit plasmid, pTM-PolI-WSN-All; 6-unit plasmid, pTM-PolI-WSN-PB2-PB1-PA-NP-M-NS; 2-unit plasmid, pTM-PolI-WSN-HA-NA; 8 × 1-unit plasmid, combination of pPolI-WSN-PB2, -PB1, -PA, -HA, -NP, -NA, -M, -NS. p.t., posttransfection.