A facile method for high-throughput co-expression of protein pairs

Mol Cell Proteomics. 2004 Sep;3(9):934-8. doi: 10.1074/mcp.T400008-MCP200. Epub 2004 Jul 7.

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

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • DNA, Complementary / genetics
  • Escherichia coli / genetics
  • Gene Expression
  • Genetic Vectors
  • Open Reading Frames
  • Plasmids / genetics
  • Proteomics / methods*
  • Protozoan Proteins / biosynthesis
  • Protozoan Proteins / genetics
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / biosynthesis
  • Saccharomyces cerevisiae Proteins / genetics

Substances

  • DNA, Complementary
  • Protozoan Proteins
  • Recombinant Proteins
  • Saccharomyces cerevisiae Proteins