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.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Base Sequence
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DNA, Complementary / genetics
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Escherichia coli / genetics
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Gene Expression
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Genetic Vectors
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Open Reading Frames
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Plasmids / genetics
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Proteomics / methods*
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Protozoan Proteins / biosynthesis
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Protozoan Proteins / genetics
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Recombinant Proteins / biosynthesis*
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Recombinant Proteins / genetics*
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Saccharomyces cerevisiae Proteins / biosynthesis
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Saccharomyces cerevisiae Proteins / genetics
Substances
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DNA, Complementary
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Protozoan Proteins
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Recombinant Proteins
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Saccharomyces cerevisiae Proteins