Fig. 7.
Southern hybridization blot of low-molecular-weight DNA from the SGs of AAVhEPO-treated animals. Vector (109 particles per animal) was administered, and Hirt-extracted DNA was prepared from SGs after 8 weeks, as described in Materials and Methods. Undigested Hirt-extracted DNA showed that the recombinant AAVhEPO viral DNA is 1,958 bp (lane a). Digestion of the Hirt-extracted DNA with SmaI (lane b) and BssHII (lane c) resulted in 1,264- and 1,740-bp fragments respectively, as expected (Fig. 1). Blue and red colors correspond to the red and blue lines in Fig. 1, indicating the restriction enzyme sites and the resulting DNA fragments. Digestion with Plasmid Safe (an enzyme only cutting linear DNA forms) indicated the presence of episomally maintained circular forms (lane d). Hirt-extracted DNA from naïve animals yielded no hybridization positive bands on Southern analyses (data not shown).