Summary
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.
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Acknowledgments
The authors thank Dr. Marianne Schüz (Mainz) for editing the manuscript. This work was supported by the Bundesministerium fü Forschung und Technologie (07QV8942) and by the Deutsche Forschungsgemeinschaft (Scha 344/1 3).
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Meffert, R., Dose, K., Rathgeber, G., Schêfer, HJ. (2009). Ultraviolet Crosslinking of DNA-Protein Complexes via 8-Azidoadenine. In: Leblanc, B., Moss, T. (eds) DNA-Protein Interactions. Methods in Molecular Biology™, vol 543. Humana Press. https://doi.org/10.1007/978-1-60327-015-1_24
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DOI: https://doi.org/10.1007/978-1-60327-015-1_24
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