Ionizing and ultraviolet radiation enhances the efficiency of DNA mediated gene transfer in vitro
The enhancement effects of ionizing and non-ionizing radiation on the efficiency of DNA mediated gene transfer were studied. Confluent Rat-2 cells were transfected with purified SV40 viral DNA, irradiated with either X-rays or ultraviolet, trypsinized, plated, and assayed for the formation of foci on Rat-2 monolayers. Both ionizing and ultraviolet radiation enhanced the frequency of A-gene transformants/survivor compared to unirradiated transfected cells. These enhancements were non-linear and dose dependent. A recombinant plasmid, pOT-TK5, was constructed that contained the SV40 virus A-gene and the Herpes Simplex virus (HSV) thymidine kinase (TK) gene. Confluent Rat-2 cells transfected with pOT-TK5 DNA and then immediately irradiated with either X-rays or 330 MeV/amu argon particles at the Berkeley Bevalac showed a higher frequency of HAT/sup +/ colonies/survivor than unirradiated transfected cells. Rat-2 cells transfected with the plasmid, pTK2, containing only the HSV TK-gene were enhanced for TK-transformation by both X-rays and ultraviolet radiation. The results demonstrate that radiation enhancement of the efficiency of DNA mediated gene transfer is not explained by increased nuclear uptake of the transfected DNA. Radiation increases the competence of the transfected cell population for genetic transformation. Three models for this increased competence are presented. The targeted integration model, the inducible recombination model, the partition model, and the utilization of DNA mediated gene transfer for DNA repair studies are discussed. 465 references.
- Research Organization:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- DOE Contract Number:
- AC03-76SF00098
- OSTI ID:
- 6209526
- Report Number(s):
- LBL-18303; ON: DE85004896
- Resource Relation:
- Other Information: Thesis
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ANIMAL CELLS
BIOLOGICAL RADIATION EFFECTS
ONCOGENIC TRANSFORMATIONS
ARGON
BIOLOGICAL MODELS
CELL CULTURES
DNA REPAIR
EXPERIMENTAL DATA
GENE RECOMBINATION
HEAVY IONS
PLASMIDS
RATS
SIMIAN VIRUS
ULTRAVIOLET RADIATION
X RADIATION
ANIMALS
BIOLOGICAL EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CELL CONSTITUENTS
CHARGED PARTICLES
DATA
ELECTROMAGNETIC RADIATION
ELEMENTS
FLUIDS
GASES
INFORMATION
IONIZING RADIATIONS
IONS
MAMMALS
MICROORGANISMS
NONMETALS
NUMERICAL DATA
PARASITES
RADIATION EFFECTS
RADIATIONS
RARE GASES
RECOVERY
REPAIR
RODENTS
VERTEBRATES
VIRUSES
560121* - Radiation Effects on Cells- External Source- (-1987)