A slot blot method for detection of ultraviolet photoproducts in DNA.
A simple new method was established to detect the major uv photoproducts in DNA. A slot blot method, involving the use of T4 DNA polymerase-associated (3'--> 5') exonuclease digestion of uv-irradiated DNA, was used to detect (primarily) cyclobutane pyrimidine dimers and pyrimidine-pyrimidone (6-4) dimers. Hydrolysis of DNA by this enzyme is quantitatively blocked by both of these photoproducts. To detect (primarily) (6-4) dimers, Escherichia coli DNA photolyase was utilized to specifically reverse cyclobutane pyrimidine dimers.
D004247:DNA / Q000737:chemistry / Q000528:radiation effects*; D004259:DNA-Directed DNA Polymerase; D004255:Deoxyribodipyrimidine Photo-Lyase; D004307:Dose-Response Relationship, Radiation; D004926:Escherichia coli / Q000201:enzymology; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases; D007202:Indicators and Reagents; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000737:chemistry / Q000528:radiation effects*; D011740:Pyrimidine Dimers / Q000032:analysis*; D015183:Restriction Mapping; D014466:Ultraviolet Rays; D014764:Viral Proteins
D007202:Indicators and Reagents; D011740:Pyrimidine Dimers; D014764:Viral Proteins; C069983:gene 43 protein, Enterobacteria phage T4; D004247:DNA; D004259:DNA-Directed DNA Polymerase; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V; D004255:Deoxyribodipyrimidine Photo-Lyase