A shuttle vector system for studying ionizing radiation-induced mutagenesis in mammalian cells

A shuttle vector system was developed to quantitate and analyze ionizing radiation-induced mutation in mammalian host cells, COS-1 and CV-1. The shuttle vector pSV2-lacY, which was constructed to detect both point mutations and deletions, was irradiated in vitro with 60Co gamma rays before introduction into unirradiated host cells. The plasmid was then isolated and reintroduced into HB101 (lacY-) bacterial host cells for identification of mutated lacY marker genes. Gamma-irradiation produced a decrease of the survival (recovery) and an increase of mutation of the shuttle vector. The mutated shuttle vector molecules were examined for structural changes by means of restriction endonuclease digestion and agarose gel electrophoresis. A dose dependent increase was observed in the percentages of gross alteration events of total mutations in mammalian host. This system will be useful for studies of ionizing radiation-induced mutagenesis.