title: A facile method for high-throughput co-expression of protein pairs.
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.
authors.affiliation: Structural Genomics of Pathogenic Protozoa (SGPP) Consortium and Center for Human Genetics and Molecular Pediatric Disease, University of Rochester Medical Center, Rochester, NY 14642, USA.
authors.forename: Andrei
authors.initials: A
authors.lastname: Alexandrov
authors.forename: Marissa
authors.initials: M
authors.lastname: Vignali
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: LaCount
authors.forename: Erin
authors.initials: E
authors.lastname: Quartley
authors.forename: Christina
authors.initials: C
authors.lastname: de Vries
authors.forename: Daniela
authors.initials: D
authors.lastname: De Rosa
authors.forename: Julie
authors.initials: J
authors.lastname: Babulski
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Mitchell
authors.forename: Lori W
authors.initials: LW
authors.lastname: Schoenfeld
authors.forename: Stanley
authors.initials: S
authors.lastname: Fields
authors.forename: Wim G
authors.initials: WG
authors.lastname: Hol
authors.forename: Mark E
authors.initials: ME
authors.lastname: Dumont
authors.forename: Eric M
authors.initials: EM
authors.lastname: Phizicky
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: Grayhack
mesh_terms: D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D016366:Open Reading Frames; D010957:Plasmids / Q000235:genetics; D040901:Proteomics / Q000379:methods*; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins