Unit

UNIT 16.8 Expression and Purification of Thioredoxin Fusion Proteins

  1. John McCoy,
  2. Edward LaVallie

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1608s28

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

McCoy, J. and LaVallie, E. 2001. Expression and Purification of Thioredoxin Fusion Proteins. Current Protocols in Molecular Biology. 28:I:16.8:16.8.1–16.8.14.

Author Information

  1. Genetics Institute, Cambridge, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1994

Abstract

This unit describes a gene fusion expression system that uses thioredoxin, the product of the Escherichia coli trxA gene, as the fusion partner. The system is particularly useful for high-level production of soluble fusion proteins in the E. coli cytoplasm; in many cases heterologous proteins produced as thioredoxin fusion proteins are correctly folded and display full biological activity. Protein fusions to His-patch Trx can usually be purified in a single step from cell lysates. Additional protocols describe E. coli cell lysis using a French pressure cell and fractionation, osmotic release of thioredoxin fusion proteins from the E. coli cytoplasm, and heat treatment to purify some thioredoxin fusion proteins.