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A facile method for high-throughput co-expression of protein pairs
Andrei Alexandrov1, Marissa Vignali, Douglas J LaCount
1Structural Genomics of Pathogenic Protozoa (SGPP) Consortium and Center for Human Genetics and Molecular Pediatric Disease, University of Rochester Medical Center, Rochester, NY 14642, USA.
Abstract:
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.