The use of antibodies to 5-bromo-2'-deoxyuridine for the isolation of DNA sequences containing excision-repair sites.
We have developed an immunological method for isolation and identification of DNA sequences containing 5-bromo-2'-deoxyuridine (BrdUrd) incorporated during UV-induced excision-repair synthesis. DNA fragments containing BrdUrd incorporated during repair synthesis were incubated with goat anti-BrdUrd and rabbit anti-goat IgG, and the antibody-DNA complexes were separated from bulk DNA by nitrocellulose filter binding. With this method, 80% of DNA sequences containing BrdUrd-labeled excision-repair sites were recovered, contaminated with less than 1% of DNA fragments devoid of excision-repair sites. Recovery of DNA fragments containing repair sites was independent of size from 2 to 20 kilobases. We have used this method in conjunction with blot hybridization to demonstrate that repair synthesis occurs in human ribosomal gene sequences in cells treated with UV.
D000906:Antibodies; D001483:Base Sequence; D001973:Bromodeoxyuridine / Q000032:analysis*; D002460:Cell Line; D004247:DNA / Q000302:isolation & purification*; D004260:DNA Repair; D004275:DNA, Ribosomal; D005347:Fibroblasts / Q000528:radiation effects; D005796:Genes; D006801:Humans; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D014466:Ultraviolet Rays
D000906:Antibodies; D004275:DNA, Ribosomal; D004247:DNA; D001973:Bromodeoxyuridine