Sequence specific double-stranded DNA detection by protein reassembly

ORGN 674

Jason R Porter, jrporter@email.arizona.edu1, Cliff I Stains, cstains@email.arizona.edu1, Aik T Ooi, atooi@ucdavis.edu2, David J Segal, djsegal@ucdavis.edu2, and Indraneel Ghosh, ghosh@email.arizona.edu1. (1) Department of Chemistry, University of Arizona, 1306 East University Blvd., Tucson, AZ 85721, (2) Department of Pharmacology, University of California, Davis, 451 E. Health Sciences Drive, Davis, CA 95616
Our lab has recently developed a novel split-protein approach for the direct detection of double-stranded DNA. Termed SEquence-Enabled Reassembly (SEER), this approach utilizes fusion proteins which combine the sequence specific nature of two zinc-finger DNA binding domains with the signal generating power of a dissected reporter protein. Recognition of a specific double-stranded DNA sequence by the two zinc-finger domains facilitates the reassembly of the associated reporter protein resulting in an observable signal. Double-stranded DNA detection using the SEER approach has been demonstrated using both the emission of light by the Green Fluorescent Protein and the colorimetric signal generated by the β-lactamase catalyzed hydrolysis of nitrocefin. Work is currently focused on expanding the chemical repertoire of SEER through the use of the bioluminescent enzyme Firefly Luciferase, which catalyzes the light emitting, ATP dependent, oxidation of D-luciferin.
 

Lipids, Nucleotides, and Mimetics
1:00 PM-4:40 PM, Wednesday, 13 September 2006 Moscone Center -- Room 131, Oral

Division of Organic Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006