Fig 5.

A transcription-deficient L mutant of RVFV. (a) trVLP production. Donor cells were transfected with VLP plasmid mixes for RVFV, using wt L or the D111A mutant of RVFV L. Omission of L constructs and envelope GPs served as controls. Supernatants containing VLPs were harvested after 72 h of transfection, and reporter activities of the cell lysates were measured. (b) Reporter activities of VLP-infected indicator cells expressing RVFV N and wt L after 24 h of incubation. Only values for the Ren-Luc minigenome reporter are shown. Reporter activities of the cotransfected FF-Luc plasmid indicated comparable transfection efficiencies (data not shown). Mean values and standard deviations from three independent experiments are shown. (c) Quantification of positive-sense RVFV minigenome RNA. Cells were transfected with plasmids of the RVFV minigenome system (with 10 ng vRen plasmid), and 72 h later, RNA levels were determined by strand-specific real-time quantitative RT-PCR (RT-qPCR) as indicated for Fig. 4. rel., relative. Mean values and standard deviations from three independent experiments are shown.