Unit

UNIT 20.3 Phage-Based Expression Cloning to Identify Interacting Proteins

  1. Julie M. Stone

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb2003s39

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Stone, J. M. 2001. Phage-Based Expression Cloning to Identify Interacting Proteins. Current Protocols in Molecular Biology. 39:20.3:20.3.1–20.3.9.

Author Information

  1. University of Missouri, Columbia, Columbia, Missouri

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUL 1997

Abstract

Phage-based expression cloning is a simple, rapid, and powerful technique to identify interacting proteins. A protein of interest is expressed as a recombinant fusion protein and labeled with 32P at an engineered recognition site to facilitate detection. b-gal proteins that are fused in-frame to cDNA inserts in a phage-derived expression library are produced by the phage and adsorbed onto nitrocellulose filters. The filters are then screened with the radioactive protein probe to identify phage clones that express the interacting protein. This technique leads directly to the isolation of a cDNA encoding the interacting protein, bypassing the need for labor-intensive protein purification, microsequencing, or antibody production.