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Measurement of DNA damage and repair in the λLIZ transgene in a Big Blue® rat cell line by quantitative PCR
Author(s) -
Yang Haiyan,
Kotturi Gopaul P.,
de Boer Johan G.,
Glickman Barry W.
Publication year - 1999
Publication title -
environmental and molecular mutagenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1
H-Index - 87
eISSN - 1098-2280
pISSN - 0893-6692
DOI - 10.1002/(sici)1098-2280(1999)33:1<21::aid-em3>3.0.co;2-1
Subject(s) - dna damage , dna repair , nucleotide excision repair , microbiology and biotechnology , transgene , biology , dna , mutagen , cell culture , genomic dna , insert (composites) , mutagenesis , genetics , gene , mutation , mechanical engineering , engineering
DNA damage and its subsequent repair occur heterogeneously throughout the genome, which reflects the nature of the damaging agents, gross chromosomal structure, the specific nucleotide sequence, and transcriptional status. We selected to investigate the repair of DNA damage in an artificial, transgenic situation. Here we report the repair of UV and BPDE‐induced DNA damage in the nontranscribed λ construct of the Big Blue® rat‐2 transgenic cell line. This was determined by quantitative PCR (QPCR) using genomic DNA isolated at specific times following treatment with UV or BPDE. The results indicate that, despite the absence of transcription, lesions induced in the lacI ‐containing λ insert by UV and BPDE are efficiently repaired. The half‐life of the polymerase‐blocking lesions is 4.2 and 5.5 hr for UV and BPDE induced lesions, respectively. This is an important observation vis‐a‐vis the use of this transgene as a model for studies of mutational mechanism. Environ. Mol. Mutagen. 33:21–27, 1999 © 1999 Wiley‐Liss, Inc.