Fig 6.

Validation of insertion profiling data. Tolerated insertion sites maintained for three passages were introduced into the pT7:MNV 3′ RZ (WT) infectious clone and yielded infectious virus by use of the DNA-based reverse genetics system. Numbers refer to the genome position after which the insertion was made. Insertions colored white lie within NS1-2, those in light gray are insertions in NS4, and those in dark gray are in VP2. Importantly, introduction of an insertion at position 6722, which was deemed replication incompetent in the profiling, failed to yield infectious virus by reverse genetics. The pT7:MNV 3′ RZ NS7 frameshift clone (FS) was used as a negative control for virus recovery. Significance was tested using one-way analysis of variance and Dunnett's multiple-comparison posttest to compare each insertion virus to the WT control, n = 3. **, P < 0.01.