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Originally published In Press as doi:10.1074/mcp.T400008-MCP200 on July 7, 2004.
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Molecular & Cellular Proteomics 3:934-938, 2004.
© 2004 by The American Society for Biochemistry and Molecular Biology, Inc.

Technology

A Facile Method for High-throughput Co-expression of Protein Pairs*,S

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

From the Structural Genomics of Pathogenic Protozoa (SGPP) Consortium and {ddagger} Center for Human Genetics and Molecular Pediatric Disease, University of Rochester Medical Center, Rochester, NY 14642; § Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, NY 14642; Departments of Genome Sciences and Medicine, University of Washington, Seattle, WA 98195; || Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195; and ** Department of Biochemistry, University of Washington, Seattle, WA 98195

We developed a method to co-express protein pairs from collections of otherwise identical Escherichia 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.

{ddagger}{ddagger} To whom correspondence should be addressed: Center for Human Genetics and Molecular Pediatric Disease, University of Rochester Medical Center, Rochester, NY 14642. Tel.: 585-275-2765; E-mail: elizabeth_grayhack{at}urmc.rochester.edu


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