Fig. 2

Diagrammatic sketch of PCR-based reverse genetics. BSR cells were seeded into 6-well tissue culture plates and when the cells reached 90% confluency, 3200 ng of seven helper plasmids were transfected into the BSR cells. The BTV proteins were expressed and necessary protein for BTV assembly and replication were prepared. Then 2000 ng of 10 PCR amplicons were transfected into BSR cells. BTV genome genes were produced by T7 polymerase in BSR cells. Then new BTV were recovered with these proteins and genome genes. In most case, the CPE could be observed 3–5 days after PCR amplicons transfection