The temperature-sensitive mutant 4 of adenovirus type 2 (Ad-2) is capable of complementing adeno-associated virus type 1 (AAV-1) in HEp2, KB and HEK cells at 34 °C and 39 °C when used as a helper virus. Heavy non-infectious AAV-1 particles can be generated by using the mutant 4 in HEp2 cells. When AAV-1 is grown in serial passages in HEp2 cells, both the wild-type Ad-2 and the mutant 4 give rise to heavy, less infectious AAV-1 particles. The heavy AAV-1 particles generated by Ad-2 in advanced serial passages retain the property of having CF and IF antigens, but the AAV-1 generated by the mutant in advanced serial passages lose this property. There is no appreciable difference in the particle counts made by electron microscopy of AAV-1 preparations generated either by Ad-2 or the mutant 4. Analysis by polyacrylamide gel electrophoresis of purified heavy AAV generated by 4 indicates that in late passage an additional polypeptide of higher mol. wt. than the three structural polypeptides is detected.


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