DNA sequence-enabled reassembly of the green fluorescent protein.
We describe a general methodology for the direct detection of DNA by the design of a split-protein system that reassembles to form an active complex only in the presence of a targeted DNA sequence. This approach, called SEquence Enabled Reassembly (SEER) of proteins, combines the ability to rationally dissect proteins to construct oligomerization-dependent protein reassembly systems and the availability of DNA binding Cys2-His2 zinc-finger motifs for the recognition of specific DNA sequences. We demonstrate the feasibility of the SEER approach utilizing the split green fluorescent protein appended to appropriate zinc fingers, such that chromophore formation is only catalyzed in the presence of DNA sequences that incorporate binding sites for both zinc fingers.
Stains
Cliff I
CI
Department of Chemistry and Department of Pharmacology and Toxicology, University of Arizona, Tucson, AZ 85721, USA.
Porter
Jason R
JR
Ooi
Aik T
AT
Segal
David J
DJ
Ghosh
Indraneel
I
D004269:DNA, Bacterial / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism*
D004269:DNA, Bacterial; D049452:Green Fluorescent Proteins