Restriction digests of the clone give the following sizes (kb): BamHI/EcoRI--4.0, 2.3, 0.5; PvuII--3.6, 1.1, 0.78, 0.6, 0.35, 0.32; XbaI--4.0, 2.8. The cotransfected cells produce a recombinant virus which can be used to infect a human cell line, allowing integration of the cloned insert into the chromosome of the cell line. Integrating vector useful for creating recombinant adeno-associated virus when cotransfected into adenovirus infected cells containing the helper virus pAAV/Ad (ATCC 68066). The depositor recommends selection of a single colony and growth in LB + amp for less than 12 hours. Because of the AAV inverted repeats, the plasmid tends to rearrange if cultivated for longer periods of time. The neoR cassette can be excised using EcoRI/BamHI or XbaI allowing replacement by a cloned gene of interest. |
