FIG. 3.

(A) DNA blot of viral DNA from AD169 and RCΔ97.08. Viral DNA was cut with PstI (lanes 1 and 3) or HindIII (lanes 2 and 4) and separated on a 0.6% agarose gel prior to transfer to a nylon membrane. The blot was hybridized with a 392-bp PCR fragment (coordinates 140479 to 140871 [Fig. 1]) that was labeled with [α-³²P]dCTP. Fragments that hybridized to the probe were visualized by autoradiography. Positions of predicted fragments are indicated by the arrows. (B) Total RNA was harvested from HEL cells infected with RCΔ97.08 or the parent virus at 48 hpi. RNA was treated with DNase and reverse transcribed by using random hexamer primers. Two adjacent amplicons were amplified by PCR, a 65-bp fragment near the translational start site and a 153-bp fragment that lies within the deleted sequences in the UL97 ORF. Amplified fragments were separated on an agarose gel, transferred to a nylon membrane, and hybridized to pON2161 random labeled with [α-³²P]dCTP. The autoradiogram is shown. Amplified products from total RNA from uninfected (lanes 1 and 4), AD169-infected (lanes 2 and 5), and RCΔ97.08-infected (lanes 3 and 6) HEL cells are shown.