Improvement of a Venus-based bimolecular fluorescence complementation assay to visualize bFos-bJun interaction in living cells

Chika Nakagawa; Kazuto Inahata; Shigenori Nishimura; Kenji Sugimoto

doi:10.1271/bbb.110189

Improvement of a Venus-based bimolecular fluorescence complementation assay to visualize bFos-bJun interaction in living cells

Biosci Biotechnol Biochem. 2011;75(7):1399-401. doi: 10.1271/bbb.110189. Epub 2011 Jul 7.

Authors

Chika Nakagawa¹, Kazuto Inahata, Shigenori Nishimura, Kenji Sugimoto

Affiliation

¹ Laboratory of Molecular Biology and Cell Informatics, Division of Bioscience and Informatics, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka, Japan. cnakagawa@bioinfo.osakafu-u.ac.jp

PMID: 21737916
DOI: 10.1271/bbb.110189

Abstract

Bimolecular fluorescence complementation (BiFC) assay makes it possible to visualize protein-protein interactions in living cells. In this assay, Venus, a bright-yellow variant of green fluorescent protein, is known to produce fluorescent backgrounds due to non-specific interactions. In this study we found that the V150A mutation increased by 8.6-fold the signal-to-noise ratio in the BiFC assay of bFos-bJun interaction.

MeSH terms

Bacterial Proteins / chemistry*
Bacterial Proteins / genetics
Biological Assay / methods
Fluorescent Dyes / chemistry*
Luminescent Proteins / chemistry*
Luminescent Proteins / genetics
Microscopy, Fluorescence / methods*
Mutagenesis, Site-Directed / methods*
Point Mutation
Protein Binding
Protein Interaction Mapping
Proto-Oncogene Proteins c-fos / analysis
Proto-Oncogene Proteins c-fos / chemistry*
Proto-Oncogene Proteins c-jun / analysis
Proto-Oncogene Proteins c-jun / chemistry*
Transfection

Substances

Bacterial Proteins
Fluorescent Dyes
Luminescent Proteins
Proto-Oncogene Proteins c-fos
Proto-Oncogene Proteins c-jun
yellow fluorescent protein, Bacteria