TABLE 1.
Optimal amounts of plasmid DNA required for the transcription and replication of an artificial Ebola virus minigenomea
| Amt of Plasmid DNA (μg) expressing:
|
Relative efficiency of GFP expressionb | |||||
|---|---|---|---|---|---|---|
| L | NP | VP35 | VP30 | T7 Pol | GFP-vRNA | |
| 1.0 | 0.5 | 0.5 | 1.0 | 1.0 | 1.0 | 1.0 |
| 1.0 | 0.5 | 0.5 | 0.3 | 1.0 | 1.0 | 2.5 |
| 1.0 | 0.5 | 0.5 | 0.1 | 1.0 | 1.0 | 3.2 |
| 2.0 | 0.5 | 0.5 | 1.0 | 1.0 | 1.0 | 1.5 |
| 2.0 | 0.5 | 0.5 | 0.3 | 1.0 | 1.0 | 7.5 |
| 2.0 | 0.5 | 0.5 | 0.1 | 1.0 | 1.0 | 3.7 |
| 3.0 | 0.5 | 0.5 | 1.0 | 1.0 | 1.0 | 2.9 |
| 3.0 | 0.5 | 0.5 | 0.3 | 1.0 | 1.0 | 8.3 |
| 3.0 | 0.5 | 0.5 | 0.1 | 1.0 | 1.0 | 7.2 |
| 4.0 | 0.5 | 0.5 | 1.0 | 1.0 | 1.0 | 0.0 |
| 4.0 | 0.5 | 0.5 | 0.3 | 1.0 | 1.0 | 13.9 |
| 4.0 | 0.5 | 0.5 | 0.1 | 1.0 | 1.0 | 2.5 |
a
293T cells were transfected with plasmids for the expression of the Ebola virus Zaire L, NP, VP35, and VP30 proteins and with p3E5EGFP and pC-T7Pol. Forty-eight hours later, the number of GFP-expressing cells was determined with a fluorescence microscope.
b
Determined by counting the number of GFP-expressing cells in five microscopic fields. The sample containing 1 μg of plasmids expressing L, VP30, p3E5EGFP, and pC-T7Pol and 0.5 μg of plasmids expressing NP and VP35 was chosen as the reference (value of 1); approximately 500 GFP-expressing cells were observed with these amounts of plasmid DNA.