ChemBioChem · 2009 · 70 citations · 45 references
Life SciencesBiotechnologyMolecular BiologyNucleic Acid BiochemistryProtein EngineeringRecombinant Dna TechnologyDna AssemblyDna ChemistryDna CrosslinkingDna LigationDna NanotechnologyDna StructureDna FunctionalizationDna NanomaterialsDna Nanostructure AssemblyDna Self-assemblyDna-based Self-assemblyDna-protein RecognitionProtein-dna InteractionsProtein-dna Interfaces
Abstract A simple modification allows DNA to be linked to recombinant proteins. DNA functionalized with three nitrilotriacetic acid groups forms coordination complexes with nickel ions and the His 6 ‐tag of the recombinant protein (here, GFP). This noncovalent linkage is reversible, site‐specific and has a high (nanomolar) affinity. magnified image We present a facile method for linking recombinant proteins to DNA. It is based on the nickel‐mediated interaction between a hexahistidine tag (His 6 ‐tag) and DNA functionalized with three nitrilotriacetic acid (NTA) groups. The resulting DNA–protein linkage is site‐specific. It can be broken quickly and controllably by the addition of a chelating agent that binds nickel. We have used this new linker to bind proteins to a variety of DNA motifs commonly used in the fabrication of nanostructures by DNA self‐assembly.
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