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. 2018 Jun 13;92(13):e00588-18. doi: 10.1128/JVI.00588-18

TABLE 1.

Screening for determination of the optimal amounts of 11 RVA T7 plasmids required for the generation of recombinant RVAa

Amt of plasmid (μg) expressingb:
 Efficiency of virus rescuec (no. positive/total no.) Virus titerd (PFU/ml) for expt:
VP1 VP2 VP3 VP4 VP6 VP7 NSP1 NSP2 NSP3 NSP4 NSP5 FAST 1 2 3
0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.2 2/3 <10 <10 0
0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0 1/3 <10 0 0
2.25 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0 0/3 0 0 0
0.75 2.25 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0 0/3 0 0 0
0.75 0.75 2.25 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0 0/3 0 0 0
0.75 0.75 0.75 2.25 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0 0/3 0 0 0
0.75 0.75 0.75 0.75 2.25 0.75 0.75 0.75 0.75 0.75 0.75 0 0/3 0 0 0
0.75 0.75 0.75 0.75 0.75 2.25 0.75 0.75 0.75 0.75 0.75 0 0/3 0 0 0
0.75 0.75 0.75 0.75 0.75 0.75 2.25 0.75 0.75 0.75 0.75 0 1/3 <10 0 0
0.75 0.75 0.75 0.75 0.75 0.75 0.75 2.25 0.75 0.75 0.75 0 3/3 6.8 × 105 4.3 × 104 5.0 × 104
0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 2.25 0.75 0.75 0 0/3 0 0 0
0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 2.25 0.75 0 0/3 0 0 0
0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 2.25 0 3/3 5.8 × 102 1.5 × 105 1.0 × 102
a

BHK/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.

b

Boldface indicates the plasmids whose amounts were 3-fold increased.

c

Efficiency of virus rescue in three experiments.

d

Virus titers were determined by plaque assay using the cultures.