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Submitted on June 14, 2004
Revised on July 7, 2004
Accepted on July 6, 2004

A facile method for high-throughput co-expression of protein pairs

Andrei Alexandrov, Marissa Vignali, Douglas J. LaCount, Erin Quartley, Christina de Vries, Daniela De Rosa, Julie Babulski, Sarah F. Mitchell, Lori W. Schoenfeld, Stanley Fields, Wim G. Hol, Mark E. Dumont, Eric M. Phizicky, and Elizabeth J. Grayhack

Biochemistry and Biophysics, University of Rochester, School of Medicine and Dentistry, Rochester, NY 14642

Corresponding Author: elizabeth_grayhack{at}urmc.rochester.edu

We developed a method to co-express protein pairs from collections of otherwise identical E. coli plasmids expressing different ORFs, by incorporating a 61-nucleotide sequence (LINK) into the plasmid to allow generation of tandem plasmids. Tandem plasmids are formed in a ligation-independent manner, propagate efficiently, and produce protein pairs in high quantities. This greatly facilitates co-expression for structural genomics projects that produce thousands of clones bearing identical origins and antibiotic markers.


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