Measurement of DNA damage and repair in the λLIZ transgene in a Big Blue® rat cell line by quantitative PCR
✍ Scribed by Haiyan Yang; Gopaul P. Kotturi; Johan G. de Boer; Barry W. Glickman
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 112 KB
- Volume
- 33
- Category
- Article
- ISSN
- 0893-6692
No coin nor oath required. For personal study only.
✦ Synopsis
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 construct of the Big Blue ா rat-2 transgenic cell line. This was determined by quantitative PCR (QPCR) using genomic DNA isolated at spe-cific times following treatment with UV or BPDE. The results indicate that, despite the absence of transcription, lesions induced in the lacI-containing 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-avis the use of this transgene as a model for studies of mutational mechanism.