Fig. 3: Nonsynonymous SNPs in the L gene facilitate the growth of BoDV-2.
A Schematic representation of the BoDV-2 L gene variants inserted into the expression plasmid and the full-length BoDV-2 cDNA plasmid. The positions of the substituted nucleotides compared to the reference sequence are indicated by red bars. The putative RdRp domain, PRNTase domain, and CTD are colored blue, green, and gray, respectively. B BoDV-2 minireplicon assay using the indicated BoDV-2 L variants and BoDV-2 N and P as helper plasmids. Values for Gluc activity were normalized to that of the LRG variant as a relative value of 1.0. Data are presented as the means ± SEM of three biologically independent experiments. One-way ANOVA and Dunnett’s multiple comparisons test were performed for statistical analysis. *p < 0.05; **p < 0.01. C, D Growth kinetics assay of rBoDV-2 variants. Vero cells infected with the indicated rBoDV-2 were cocultured with uninfected Vero cells at a ratio of 1 to 19 and passaged every 3 or 4 days. DPC, days post coculture. C The percentage of infected cells was determined by IFA using an antibody against BoDV N. D The copy number of viral RNA was measured via RT-qPCR using total RNA extracted from each cells. Data are presented as the means ± SEM of three biologically independent experiments. Two-way ANOVA and Tukey’s multiple comparisons test were performed for statistical analysis. Different characters (a, b, c) represent statistically significant differences at p < 0.05; ns, not significant.
