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. 1990 Nov 25;18(22):6633–6636. doi: 10.1093/nar/18.22.6633

UV-induced cross-linking of Tet repressor to DNA containing tet operator sequences and 8-azidoadenines.

R Meffert 1, G Rathgeber 1, H J Schäfer 1, K Dose 1
PMCID: PMC332621  PMID: 2251123

Abstract

The synthesis of 8-azido-2'-deoxyadenosine-5'-triphosphate is described. The photoreactive dATP analog was characterized by thin layer chromatography, proton resonance spectroscopy, infrared spectroscopy and UV spectroscopy. Its photolysis upon UV irradiation was studied. After incorporation of this dATP analog into DNA containing the tet operator sequence the investigation of the interactions between tet operator DNA and Tet repressor protein by UV photocross-linking becomes possible. Photocross-linking of protein to DNA was demonstrated by the reduced migration of the DNA in SDS polyacrylamide gel electrophoresis. Addition of the inducer tetracycline prior to UV irradiation significantly reduces the DNA-protein cross-linking rate. The long wave UV light applied here does not significantly alter the DNA or the protein under the photocross-linking conditions.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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