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A Facile Method for Reversibly Linking a Recombinant Protein to DNA
Author(s) -
Goodman Russell P.,
Erben Christoph M.,
Malo Jonathan,
Ho Wei M.,
McKee Mireya L.,
Kapanidis Achillefs N.,
Turberfield Andrew J.
Publication year - 2009
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200900165
Subject(s) - nitrilotriacetic acid , recombinant dna , linker , dna , chemistry , combinatorial chemistry , chelation , biochemistry , nanotechnology , materials science , gene , organic chemistry , computer science , operating system
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.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.