A facile method for high-throughput co-expression of protein pairs.
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.
Alexandrov
Andrei
A
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.
Vignali
Marissa
M
LaCount
Douglas J
DJ
Quartley
Erin
E
de Vries
Christina
C
De Rosa
Daniela
D
Babulski
Julie
J
Mitchell
Sarah F
SF
Schoenfeld
Lori W
LW
Fields
Stanley
S
Hol
Wim G
WG
Dumont
Mark E
ME
Phizicky
Eric M
EM
Grayhack
Elizabeth J
EJ
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
D018076:DNA, Complementary; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins