Microplate enzyme-linked immunosorbent assay for the detection of primary DNA alterations based on the interaction with UvrA/UvrB.
An enzyme-linked microplate immunoassay for the analysis of primary DNA lesions is described. The assay principle is based on the interaction of the bacterial DNA repair proteins UvrA and UvrB with DNA and on the immunodetection of UvrB forming a stable complex with covalently modified nucleotides. Using this technique we were able to detect damages in genomic DNA induced by uv light and by several different genotoxic agents. The detection sensitivity of the method reaches down to the nanomolar range of the mutagenic compound depending on the type of the DNA alteration. The method might be used in automated high-throughput studies.
Böse
G
G
Max-Planck-Institut für molekulare Physiologie, Dortmund, D-44227, Germany.
Kuhlmann
J
J
Plass
M
M
Müller
O
O
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D002417:Cattle; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D004249:DNA Damage; D004265:DNA Helicases / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins; D051379:Mice; D009994:Osmolar Concentration; D014407:Tumor Cells, Cultured
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C058093:UvrB protein, E coli; D004247:DNA; C068631:UvrA protein, E coli; D000251:Adenosine Triphosphatases; D004265:DNA Helicases