FIG 2.

Examination of revertant reporter viruses after 10 passages. (A and B) Schematic images of reporter genes before and after serial passage. Nucleotide sequences of dsRNA genomes purified from rsSA11-NLuc-Full (P10) and rsSA11-ZsG-Full (P10), as indicated, were analyzed. (C) Expression of NSP1 from reporter SA11 virus-infected cells (P1 or P10). MA104 cells were infected with rsSA11 or rsSA11-NLuc-Full (P1 and P10) or rsSA11-ZsG-Full (P1 and P10) at an MOI of 1 PFU/cell. NSP1 and VP7 in cell lysates were detected using a rabbit anti-NSP1 antibody and a monoclonal antibody specific for VP7, followed by appropriate HRP-conjugated secondary antibodies. An anti-β-actin antibody was used as a loading control. (D) Replication of rsSA11-NLuc and rsSA11-ZsG P1 (P5 and P10) viruses. MA104 cells were infected with each reporter virus at an MOI of 0.01 PFU per cell and incubated for 48 h. Infectious virus titers were measured in a plaque assay. Data are expressed as the means ± standard deviations (n = 3). *, P < 0.05 (Tukey’s multiple-comparison test).