Recombinant Lloviu virus as a tool to study viral replication and host responses
Fig 1
Complementation assays identify sequences that facilitate LLOV transcription and replication.
(A) Schematic of LLOV minigenomes with EBOV noncoding region (negative sense L gene 3ˈ UTR) and trailer (top) or hybrid 5ˈ genome ends consisting of the LLOV L gene 3ˈ UTR and trailer complemented with short terminal sequences from the EBOV trailer, designated LEX where X represents the number of added terminal nucleotides from EBOV. Le, leader; NP, nucleoprotein; L, polymerase; Tr, trailer. (B) Luciferase-based minigenome assays comparing the 3L5E [9] and chimeric 3L5LEX minigenomes. Mean fold induction of firefly luciferase activity over the negative control (minus L) and normalized to ß-galactosidase activity (transfection control) with standard deviation is shown. (C) Schematic of bicistronic 3L5LE72 minigenomes containing firefly and renilla luciferase reporters separated by either the wild-type LLOV VP24-L intergenic region (IRwt) or the same intergenic region with an inserted gene border (IRins) consisting of overlapping LLOV gene end (GE, red bar) and gene start (GS, green triangle) signals. (D) Luciferase-based minigenome assays comparing the bicistronic minigenomes. Mean fold induction of firefly and renilla luciferase activities over the negative controls (minus L) and normalized to ß-galactosidase activity (transfection control) with standard deviation is shown.