Schematic presentation of the baculovirus expression strategies used to generate RV-VLPs.

The donor plasmids contains ORFs coding for specific rotavirus proteins (labelled downstream to the promoters regulating their expression as described in the text) that were transposed into bacmids which were subsequently used to generate baculoviruses. The restriction sites used for construction of the recombinant transfer plasmids are not indicated on the pFBq plasmids maps above, see Fig. S1 and Table 1 for more details. (I) pFBq plasmid construct used to generate recombinant dualcistronic baculoviruses that was used to prepare dRV-VLPs (VP2/6) through single infection of insect cells. (II) pFBq plasmid constructs used to generate recombinant dualcistronic and monocistronic baculoviruses that were used to prepare tRV-VLPs (VP2/6/7) through simultaneous infection of insect cells. (III) pFBq plasmid constructs used to generate recombinant dualcistronic and monocistronic baculoviruses that were used to prepare tRV-VLPs (VP2/6/4) through simultaneous infection of insect cells. (IV) pFBq plasmid constructs used to generate recombinant dualcistronic baculoviruses that were used to prepare tRV-VLPs (VP2/6/7/4) through simultaneous infection of insect cells. (V) pFBq plasmid constructs used to generate recombinant dualcistronic and monocistronic baculoviruses that were used to prepare tRV-VLPs (VP2/6/7/4) through step-wise co-infection strategy. Insect cells were initially infected with dualcistronic baculoviruses confirmed to express VP2/6 and recombinant monocistronic baculoviruses confirmed to express VP4. This was followed by infection with recombinant monocistronic baculoviruses confirmed to express VP7 12 hours post initial infection (hpi).