Unit

UNIT 5.21 Recombinant Protein Complex Expression in E. coli

  1. William Selleck,
  2. Song Tan

Published Online: 1 MAY 2008

DOI: 10.1002/0471140864.ps0521s52

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Selleck, W. and Tan, S. 2008. Recombinant Protein Complex Expression in E. coli. Current Protocols in Protein Science. 52:5.21:5.21.1–5.21.21.

Author Information

  1. Center for Gene Regulation, Department of Biochemistry & Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania

Publication History

  1. Published Online: 1 MAY 2008
  2. Published Print: MAY 2008

Abstract

This unit provides procedures to design, create, and utilize polycistronic plasmids that express multicomponent protein complexes in E. coli. Both the original pST39 polycistronic expression system, which permits four genes to be coexpressed from a single plasmid, and the more recent pST44 polycistronic system, which facilitates incorporation of affinity tags and simplifies the construction of variant deletion or point mutation polycistronic plasmids, are described. Emphasis is placed on practical details for creating polycistronic expression plasmids, expressing the protein complex in E. coli, purifying the protein complex, and troubleshooting potential expression problems. Curr. Protoc. Protein Sci. 52:5.21.1-5.21.21. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • multisubunit protein complexes;
  • polycistronic expression;
  • affinity tags