Abstract |
Fibroblasts from a patient with xeroderma pigmentosum complementation group D were treated with Simian virus 40 to establish a transformed cell line suitable for studies of DNA-mediated gene transfer. After progressing through 2 crises, a stable line, XP6Be(SV40), was established and cultured for more than 1 year. This line retains the characteristic xeroderma pigmentosum ultraviolet hypersensitivity and is able to complement a SV40-transformed group A line when fused and assayed for ultraviolet radiation inhibition of colony-forming ability. XP6Be(SV40) expressed high levels of transfected chloramphenicol acetyltransferase activity (0.1 nmole X mg-1 X min-1) in a transient expression assay, showed stable expression of transfected gpt or neo genes (frequency 1-20 X 10(-5)), and permitted replication of the mutagenesis shuttle vector plasmid, pZ189. Ultraviolet treatment (500 J X m-2) of pZ189 prior to replication in XP6Be(SV40) resulted in a large reduction in plasmid yield (5% survival) and a 60-fold increase in the mutation frequency, reflecting the reduced ability of these cells to repair ultraviolet-damaged transfecting DNA. This cell line provides the opportunity to utilize transfection studies in cells with the xeroderma pigmentosum group D defect in excision repair.
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Authors | M Protić-Sabljić, S Seetharam, M M Seidman, K H Kraemer |
Journal | Mutation research
(Mutat Res)
Vol. 166
Issue 3
Pg. 287-94
(Nov 1986)
ISSN: 0027-5107 [Print] Netherlands |
PMID | 3023995
(Publication Type: Journal Article)
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Chemical References |
- Bacterial Proteins
- Recombinant Fusion Proteins
- Acetyltransferases
- Chloramphenicol O-Acetyltransferase
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Topics |
- Acetyltransferases
(biosynthesis)
- Bacterial Proteins
(biosynthesis)
- Cell Line
- Cell Transformation, Viral
- Chloramphenicol O-Acetyltransferase
- DNA Repair
- Fibroblasts
(radiation effects)
- Genetic Complementation Test
- Genetic Vectors
- Humans
- Mutation
- Plasmids
- Radiation Tolerance
- Recombinant Fusion Proteins
(biosynthesis)
- Simian virus 40
(physiology)
- Transfection
- Ultraviolet Rays
- Xeroderma Pigmentosum
(genetics, pathology)
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