UNIT 20.3 Phage-Based Expression Cloning to Identify Interacting Proteins
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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. 39:20.3.1–20.3.9.
- Published Online: 1 MAY 2001
- Published Print: JUL 1997
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.