Unit

UNIT 20.9 Quantitative Analysis of Protein-Protein Interactions by Split Firefly Luciferase Complementation in Plant Protoplasts

  1. Jian-Feng Li,
  2. Dandan Zhang

Published Online: 1 JUL 2014

DOI: 10.1002/0471142727.mb2009s107

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Li, J.-F. and Zhang, D. 2014. Quantitative Analysis of Protein-Protein Interactions by Split Firefly Luciferase Complementation in Plant Protoplasts. Current Protocols in Molecular Biology. 107:20.9:20.9.1–20.9.8.

Author Information

  1. Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts

Publication History

  1. Published Online: 1 JUL 2014

Abstract

This unit describes the split firefly luciferase complementation (SFLC) assay, a high-throughput quantitative method that can be used to investigate protein-protein interactions (PPIs) in plant mesophyll protoplasts. In SFLC, the two proteins to be tested for interaction are expressed as chimeric proteins, each fused to a different half of firefly luciferase. If the proteins interact, a functional luciferase can be transitorily reconstituted, and is detected using the cell-permeable substrate d-luciferin. An advantage of the SFLC assay is that dynamic changes in PPIs in a cell can be detected in a near real-time manner. Another advantage is the unusually high DNA co-transfection and protein expression efficiencies that can be achieved in plant protoplasts, thereby enhancing the throughput of the method. Curr. Protoc. Mol. Biol. 107:20.9.1-20.9.8. © 2014 by John Wiley & Sons, Inc.

Keywords:

  • protein-protein interaction;
  • plant protoplasts;
  • split firefly luciferase complementation