abstract |
Described is the construction of an adeno-associated virus (AAV) plasmid containing an amber mutation within the AAV rep gene and the propagation of this mutant AAV on monkey cell lines containing an amber suppressor. It is shown that pure populations of AAV particles containing the mutant genome could be generated. The spontaneous reversion frequency of the amber mutation to wild-type was less than 10-5. This reversion frequency was significantly elevated by growth of the mutant virus in cultured human cells treated with mutagens. The amber mutant AAV particles provide a novel and sensitive assay for mutagenic agents in cultured human cells. |