TABLE 3.

Efficiency of generation of recombinant RVAs from transfected BHK/T7-9 cells^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
0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0/2	0	0
0.75	0.75	0.75	0.75	0.75	0.75	0.75	2.25	0.75	0.75	2.25	1/2	<10	0
0.75	0.75	0.75	0.75	0.75	0.75	0.75	3.75	0.75	0.75	3.75	2/2	<10	1.2 × 102

^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).

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

^cEfficiency of virus rescue in two experiments.

^dVirus titers were determined by plaque assay using the cultures of transfected BHK/T7-9 cells.