C. Muraiso et al., A SHUTTLE VECTOR SYSTEM FOR STUDYING IONIZING RADIATION-INDUCED MUTAGENESIS IN MAMMALIAN-CELLS, Journal of radiation research, 34(2), 1993, pp. 148-156
A shuttle vector system was developed to quantitate and analyze ionizi
ng radiation-induced mutation in mammalian host cells, COS-1 and CV-1.
The shuttle vector pSV2-lacY, which was constructed to detect both po
int mutations and deletions, was irradiated in vitro with Co-60 gamma
rays before introduction into unirradiated host cells. The plasmid was
then isolated and reintroduced into HB101 (lacY-) bacterial host cell
s for identification of mutated lacY marker genes. Gamma-irradiation p
roduced a decrease of the survival (recovery) and an increase of mutat
ion of the shuttle vector. The mutated shuttle vector molecules were e
xamined for structural changes by means of restriction endonuclease di
gestion and agarose gel electrophoresis. A dose dependent increase was
observed in the percentages of gross alteration events of total mutat
ions in mammalian host. This system will be useful for studies of ioni
zing radiation-induced mutagenesis.