Unit

UNIT 12.9 Analysis of DNA-Protein Interactions Using Proteins Synthesized In Vitro from Cloned Genes

  1. Kevin Struhl

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1209s24

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Struhl, K. 2001. Analysis of DNA-Protein Interactions Using Proteins Synthesized In Vitro from Cloned Genes. Current Protocols in Molecular Biology. 24:12.9:12.9.1–12.9.4.

Author Information

  1. Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1993

Abstract

To detect DNA binding activity, radiolabeled protein is incubated with specific DNA fragments, and protein-DNA complexes are separated from free protein by electrophoresis in native acrylamide gels. Unlike the more conventional mobility shift assay which utilizes 32P-labeled DNA and unlabeled protein, the assay described here generally utilizes 35S-labeled protein and unlabeled DNA. Major advantages of this method are that any desired mutant protein can be tested for its DNA-binding properties simply by altering the DNA template, and the subunit structure (e.g., dimer, tetramer) can be determined.