TABLE 2.

Determination of the synergistic interaction between the two plasmids encoding the NSP2 and NSP5 genes on the generation of recombinant RVA^a

Amt of rescue T7 plasmid (μg) expressingb:											Efficiency of virus rescuec (no. positive/total no.)	Virus titerd (PFU/ml) in expt:
VP1	VP2	VP3	VP4	VP6	VP7	NSP1	NSP2	NSP3	NSP4	NSP5		1	2	3	4	5	6
0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	2/6	<10	25	0	0	0	0
0.75	0.75	0.75	0.75	0.75	0.75	0.75	2.25	0.75	0.75	0.75	6/6	2.0 × 104	<10	3.8 × 104	4.8 × 103	2.8 × 104	5.0 × 104
0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	2.25	4/6	1.5 × 105	27.5	6.0 × 103	15	0	0
0.75	0.75	0.75	0.75	0.75	0.75	0.75	2.25	0.75	0.75	2.25	6/6	3.3 × 106	6.5 × 105	2.5 × 104	3.3 × 105	1.0 × 106	7.5 × 105
0.75	0.75	0.75	0.75	0.75	0.75	0.75	3.75	0.75	0.75	0.75	6/6	2.5 × 104	2.8 × 105	2.8 × 104	4.5 × 104	5.0 × 104	3.0 × 105
0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	3.75	5/6	45	0	75	4.0 × 104	6.0 × 102	42.5
0.75	0.75	0.75	0.75	0.75	0.75	0.75	3.75	0.75	0.75	3.75	6/6	7.5 × 105	8.0 × 104	1.2 × 106	5.3 × 105	6.5 × 104	2.8 × 105

^aBHK/T7-9 cells in 6-well plates were transfected with the indicated amounts of plasmids. One day later, the transfected cells were washed with incomplete medium and then cultured for 3 days in incomplete medium containing trypsin (0.3 μg/ml) with overlaid CV-1 cells.

^bBoldface indicates the plasmids whose amounts were increased (3- or 5-fold).

^cEfficiency of virus rescue in six experiments.

^dVirus titers were determined by plaque assay using the cultures.