Summmary
Visualization of protein–protein interactions in vivo offers a powerful tool to resolve spatial and temporal aspects of cellular functions. Bimolecular fluorescence complementation (BiFC) makes use of nonfluorescent fragments of green fluorescent protein or its variants that are added as “tags” to target proteins under study. Only upon target protein interaction is a fluorescent protein complex assembled and the site of interaction can be monitored by microscopy. In this chapter, we describe the method and tools for use of BiFC in the yeast Saccharomyces cerevisiae.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Tsien, R. Y. (1998) The green fluorescent protein. Annu. Rev. Biochem. 67, 509–544.
Hu, C-D. Chinenov, Y., and Kerppola, T. K. (2002) Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation. Mol Cell. 9, 789–798.
Miyawaki A. (2003) Visualization of the spatial and temporal dynamics of intracellular signaling. Dev Cell. 4, 295–305.
Tsien, R. Y. Bacskai, B. J., and Adams, S. R. (1993) FRET for studying intracellular signalling. Trends Cell Biol. 3, 242–245.
Kerppola, T. K. (2006) Visualization of molecular interactions by fluorescence complementation. Nat Rev Mol Cell Biol. 7, 449–456.
Hu, C-D. Grinberg, A., and Kerppola, T. (2005) Visualization of protein interaction in living cells using bimolecular fluorescence complementation (BiFC) analysis. In: Current Protocols in Cell Biology. (Bonifacino JS, Dasso M, Harford JB, Lippincott-Schwartz J, Yamada KM., ed). Wiley, Hoboken, NJ: pp. 21.3.1–21.3.21.
Shyu, Y.J., Liu, H., Deng, X., and Hu, C-D. (2006) Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis under physiological conditions. Biotechniques. 40, 61–66.
Jach, G., Pesch, M., Richter, K., Frings, S., and Uhrig, J.F. (2006) An improved mRFP1 adds red to bimolecular fluorescence complementation. Nat, Methods. 3, 597–600.
Hu, C-D., and Kerppola, T. K. (2003) Simultaneous visualization of multiple protein interactions in living cells using multicolor fluorescence complementation analysis. Nat. Biotechnol. 21, 539–545.
Mumberg, D., Muller, R., and Funk, M. (1995) Yeast vectors for the controlled expression of heterologous proteins in different genetic backgrounds. Gene. 156, 119–122.
Sherman, F. (1983) Getting started with yeast. In: Methods in enzymology (Fink, G. and Hicks, J. B., eds). Academic Press, New York, NY: pp. 3–20.
Acknowledgments
Chang-Deng Hu is acknowledged for EYFP fragment-containing plasmids and comments on the manuscript. Marko Kaksonen is thanked for helpful comments on live cell imaging. The work was supported by Academy of Finland grant No. 211171 to J.J.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Skarp, KP., Zhao, X., Weber, M., Jäntti, J. (2008). Use of Bimolecular Fluorescence Complementation in Yeast Saccharomyces cerevisiae . In: Vancura, A. (eds) Membrane Trafficking. Methods in Molecular Biology, vol 457. Humana Press. https://doi.org/10.1007/978-1-59745-261-8_12
Download citation
DOI: https://doi.org/10.1007/978-1-59745-261-8_12
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-925-3
Online ISBN: 978-1-59745-261-8
eBook Packages: Springer Protocols