FIG 2.
Design and evaluation of multicistronic minigenomes. (A) Schematic overview of the different mono- and multicistronic minigenomes used in this study. Open reading frames encoding Renilla luciferase (Rluc) and the viral proteins VP40, GP1,2, and VP24 are shown as light boxes, and noncoding regions (NCR), including the terminal leader and trailer regions, are shown as dark boxes, with the subscript indicating which NCR was used to join the open reading frames. All minigenomes used in this study are negative-sense (vRNA) minigenomes. (B) Influence of minigenome length on reporter activity. Minigenome assays were performed in 293 cells using the different mono- and multicistronic minigenomes depicted in panel A, using the same molar amounts of minigenome plasmids for transfection. Reporter activity (in relative light units) was measured 3 days after transfection. The means and standard deviations from 6 independent experiments are shown, as are a linear regression curve and its coefficient of correlation (R2).