Figure 2.

Linear episomal forms of rAAV2 persist in the skeletal muscle of SCID (DNA-PKcs-deficient) mice. Samples (30 μg/lane) of muscle DNA from vector injection sites were incubated without restriction enzyme (−) or with XmnI (X, no cutting site within the vector), HindIII (H, cuts the vector once), BglII (B, cuts the vector twice), or SacI (S, cuts the vector twice). Southern hybridizations were performed with ³²P-labeled probes derived from either the hAAT cDNA (AAT) or from the CMV promoter (CMV) as labeled. A shows results from the rAAV vector (C-AT)-injected and the PBS-injected SCID mice killed 18 weeks after injection. To the left of the autoradiograph are diagrams of predicted HindIII (H) and BglII (B) digestion patterns of the vector DNA in the free ends and all possible junctions, including head-to-head (H-H), tail-to-tail (T-T), and head-to-tail (H-T) junctions. B shows the DNA samples (60 μg/lane) from mice killed 52 weeks after injection. To the left of the autoradiograph are diagrams of predicted HindIII (H) and ScaI (S) digestion patterns.