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Title: Ionizing and ultraviolet radiation enhances the efficiency of DNA mediated gene transfer in vitro

Abstract

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 recombinationmore » model, the partition model, and the utilization of DNA mediated gene transfer for DNA repair studies are discussed. 465 references.« less

Authors:
Publication Date:
Research Org.:
Lawrence Berkeley Lab., CA (USA)
OSTI Identifier:
6209526
Report Number(s):
LBL-18303
ON: DE85004896
DOE Contract Number:
AC03-76SF00098
Resource Type:
Technical Report
Resource Relation:
Other Information: Thesis
Country of Publication:
United States
Language:
English
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; 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)

Citation Formats

Perez, C.F. Ionizing and ultraviolet radiation enhances the efficiency of DNA mediated gene transfer in vitro. United States: N. p., 1984. Web. doi:10.2172/6209526.
Perez, C.F. Ionizing and ultraviolet radiation enhances the efficiency of DNA mediated gene transfer in vitro. United States. doi:10.2172/6209526.
Perez, C.F. Wed . "Ionizing and ultraviolet radiation enhances the efficiency of DNA mediated gene transfer in vitro". United States. doi:10.2172/6209526. https://www.osti.gov/servlets/purl/6209526.
@article{osti_6209526,
title = {Ionizing and ultraviolet radiation enhances the efficiency of DNA mediated gene transfer in vitro},
author = {Perez, C.F.},
abstractNote = {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.},
doi = {10.2172/6209526},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Wed Aug 01 00:00:00 EDT 1984},
month = {Wed Aug 01 00:00:00 EDT 1984}
}

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