Ultraviolet crosslinking of DNA protein complexes via 8-azidoadenine.
The synthesis of 8-azido-2'-deoxyadenosine-5'-triphosphate is described. The photoreactive dATP analog was characterized by thin layer chromatography and UV spectroscopy. Its photoreactivity upon UV irradiation was studied. After incorporation of this dATP analog by nick translation into DNA containing the tet operator sequence the investigation of the interactions between tet operator DNA and Tet repressor becomes possible. Photocrosslinking of protein to DNA was demonstrated by the reduced migration of the DNA protein crosslinks in SDS polyacrylamide gel electrophoresis.
Meffert
Rainer
R
Johannes Gutenberg-Universitêt, Fachbereich Chemie, Pharmazie und Geowissenschaften, Institut fü Biochemie, 55099 Mainz, Germany.
Dose
Klaus
K
Rathgeber
Gabriele
G
Schêfer
Hans-Jochen
HJ
D000225:Adenine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000528:radiation effects; D000227:Adenine Nucleotides / Q000737:chemistry; D001386:Azides / Q000737:chemistry* / Q000528:radiation effects*; D002852:Chromatography, Ion Exchange; D003432:Cross-Linking Reagents / Q000737:chemistry* / Q000528:radiation effects*; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D018736:DNA Adducts / Q000378:metabolism; D003838:Deoxyadenine Nucleotides / Q000138:chemical synthesis / Q000737:chemistry; D007097:Imines / Q000737:chemistry; D008967:Molecular Biology; D008968:Molecular Conformation; D019920:Photoaffinity Labels; D010957:Plasmids / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D013057:Spectrum Analysis; D014466:Ultraviolet Rays
D000227:Adenine Nucleotides; D001386:Azides; D003432:Cross-Linking Reagents; D018736:DNA Adducts; D003838:Deoxyadenine Nucleotides; D007097:Imines; D019920:Photoaffinity Labels; D011506:Proteins; C057680:8-azido-2'-deoxyadenosine-5'-triphosphate; C017621:phenylnitrene; C032235:8-azidoadenine; D004247:DNA; D000225:Adenine