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SINGLET OXYGEN INDUCED DNA DAMAGE AND MUTAGENICITY IN A SINGLE‐STRANDED SV40‐BASED SHUTTLE VECTOR
Author(s) -
Ribeiro D. T.,
Madzak C.,
Sarasin A.,
Mascio P. DI,
Sies H.,
Menck C. F. M.
Publication year - 1992
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1992.tb04207.x
Subject(s) - dna , shuttle vector , singlet oxygen , extrachromosomal dna , plasmid , mutagenesis , chemistry , dna damage , transfection , microbiology and biotechnology , guanine , phosphodiester bond , vector (molecular biology) , mutation , biophysics , biology , biochemistry , recombinant dna , oxygen , nucleotide , rna , gene , organic chemistry
— The effects of singlet oxygen ( 1 O 2 ), generated by the thermal decomposition of water soluble NDPO 2 (endoperoxide of the disodium 3,3'‐(1,4‐naphthylidene) dipropionate), on a single‐stranded shuttle vector were analysed. 1 O 2 induces a much higher level of breaks in the phosphodiester backbone of single‐stranded than double‐stranded DNA. This may be due to a higher accessibility of guanine residue, primarily damaged by 1 O 2 . The damaged vector was transfected into monkey COS7 cells where single‐stranded DNA was converted to the double‐stranded replicative form DNA. After 3 days, extrachromosomal DNA was extracted and the plasmids rescued in E. coli to study mutagenesis. There is a significant increase in mutation frequency of damaged single‐stranded DNA in comparison to untreated DNA. It is concluded that 1 O 2 induces breaks in the backbone of single‐stranded DNA and that the 1 O 2 ‐damaged molecules are mutated after passage through mammalian cells.