6533b873fe1ef96bd12d4e45
RESEARCH PRODUCT
UV-induced cross-linking of proteins to plasmid pBR322 containing 8-azidoadenine 2′-deoxyribonucleotides
Rainer MeffertKlaus Dosesubject
Ultraviolet RaysDNA polymeraseDNA polymerase IIUltraviolet irradiationBiophysicsAzidoadeninePlasmid pBR322BiochemistryHistonesDeoxyadenine NucleotidesPlasmidStructural BiologyEscherichia coliGeneticsNick translationMolecular BiologyPlasmid preparationDNA clampNick-translationbiologyDNA-protein cross-linkCell BiologyDNA Polymerase IPBR322Cross-Linking ReagentsBiochemistrybiology.proteinDNA polymerase IPlasmidsdescription
Abstract An efficient method of cross-linking DNA to protein is described. The method is based on the incorporation of photoactive 8-azidoadenine 2′-deoxyribonucleotides into DNA. We have found that 8-N 3 dATP is a substrate for E. coli DNA polymerase I and that 8-N 3 dATP can be incorporated into plasmid pBR322 by nick-translation. Subsequently we were able to cross-link a set of different proteins to 8-azido-2′-deoxyadenosine-containing pBR322 by UV irradiation (366 nm). No DNA-protein photocross-linking was observed under the same conditions when the non-photoactive plasmid pBR322 was used.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1988-11-07 | FEBS Letters |