Site-specific covalent attachment of DNA to proteins using a photoactivatable Tus-Tev complex

摘要

Investigations into the photocrosslinking kinetics of the protein Tus with various bromodeoxyuridine-substituted Ter DNA variants highlight the potential use of this complex as a photoactivatable connector between proteins of interest and specific DNA sequences.rnArtificial DNA-protein conjugates have recently attracted attention as tools for nanobiotechnology and bioanalytical chemistry. Their utility as biosensors, artificial nucleases or in protein microarrays has been successfully demonstrated. Various systems for the covalent attachment of proteins to nucleic acids have been described, including chemical crosslinking of oligonucleotides to protein lysine or cysteine residues, expressed protein ligation, chemoenzymatic reactions and the use of photoaptamers. The objective of this work was to develop a straightforward and reliable method to covalently and regiospecifically link any protein of interest (POI) with any specific DNA sequence. To do so, we made use of the DNA-binding properties of the 36 kDa Escherichia coli replication terminator protein Tus.