A Repair Competition Assay to Assess Recognition by Human Nucleotide Excision Repair | Zendy

Martin Heß | Zendy; Daniela Gunz | Zendy; Hanspeter Naegeli | Zendy

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A Repair Competition Assay to Assess Recognition by Human Nucleotide Excision Repair

Author(s) -

Martin Heß,

Daniela Gunz,

Hanspeter Naegeli

Publication year - 1996

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/24.5.824

Subject(s) - nucleotide excision repair , biology , dna , dna repair , dna damage , microbiology and biotechnology , nucleotide , genetics , biochemistry , gene

We developed a competition assay to compare, in a quantitative manner, the ability of human nucleotide excision repair (NER) to recognise structurally different forms of DNA damage. This assay uses a NER substrate consisting of M13 double-stranded DNA with a single and uniquely located acetylaminofluorene (AAF) adduct, and measures the efficiency by which multiply damaged plasmid DNA competes for excision repair of the site-directed modification. To validate this assay, we tested competitor DNA containing defined numbers of either AAF adducts or UV radiation products. In both cases, repair of the site-directed NER substrate was inhibited in a damage-specific and dose-dependent manner. We then exploited this competition assay to determine the susceptibility of bulky adozelesin-DNA adducts to human NER.

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