FIG 1.

Generation of reporter MRV. (A) Schematic showing generation of rsT1L-NLuc using an MRV reverse genetics system. The NLuc-P2A cassette was inserted into the L1 gene between nucleotides 124 and 125. To express NLuc-P2A as a fusion peptide with the L1 ORF N-terminal region, additional CC nucleotides were inserted before the NLuc-P2A cassette. The region comprising nucleotides 19 to 124 of the L1 gene was duplicated after the NLuc-P2A cassette to reconstruct the L1 ORF. (B) Electrophoresis of dsRNA purified from rsT1L and rsT1L-NLuc. Arrowheads indicate the L1wt and L1-NLuc gene segments. (C and D) Kinetics of virus replication and NLuc activity of rsT1L-NLuc in L929 cells. (E to G) Correlation between viral replication and NLuc activity of rsT1L-NLuc in human and murine cancer cell lines. Cells were infected with rsT1L-NLuc at an MOI of 0.01 PFU/cell and incubated for 48 h. Viral infectious titers and NLuc activity in the cell lysates were then determined.