FIG 5.

Site-specific mutations introduced into the genomes of the rescued VP4 gene transfectants. The VP4 gene of each virus was amplified by RT-PCR, using the primers shown in Table 2, to yield a 686-bp product. (A) The VP4 gene was amplified by use of RRV VP4-specific primers (lanes 1 to 3) and rotavirus VP4 degenerate primers (lanes 4 to 6). (B) The amplified fragments were digested with XbaI, followed by separation in a 2% agarose gel. Uncut WT VP4 (RRV) and VP4^XbaI (RRV) were run in a 2% agarose gel (lanes 1 and 2), along with WT VP4 (RRV) and VP4^XbaI (RRV) after digestion with XbaI (lanes 3 and 4).