Abstract
Bimolecular fluorescence complementation (BiFC) is a powerful method for studying protein-protein interactions in different cell types and organisms. This method was recently developed in the fruit fly Drosophila melanogaster, allowing analyzing protein interaction properties in a physiologically relevant developing context. Here we present a detailed protocol for performing BiFC with the Venus fluorescent protein in live Drosophila embryos, taking the Hox-PBC partnership as an illustrative test case. This protocol applies to any transcription factor and split fluorescent protein in general.
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
Kerppola TK (2008) Bimolecular fluorescence complementation (BiFC) analysis as a probe of protein interactions in living cells. Annu Rev Biophys 37:465–487
Kodama Y, Hu CD (2012) Bimolecular fluorescence complementation (BiFC): a 5-year update and future perspectives. Biotechniques 53:285–298
Hu CD, Chinenov Y, Kerppola TK (2002) Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation. Mol Cell 9:789–798
Shaner NC, Lambert GG, Chammas A, Ni Y, Cranfill PJ et al (2013) A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum. Nat Methods 10:407–409
Zhou J, Lin J, Zhou C, Deng X, Xia B (2011) An improved bimolecular fluorescence complementation tool based on superfolder green fluorescent protein. Acta Biochim Biophys Sin (Shanghai) 43:239–244
Kim J, Zhao T, Petralia RS, Yu Y, Peng H et al (2012) mGRASP enables mapping mammalian synaptic connectivity with light microscopy. Nat Methods 9:96–102
Shyu YJ, Liu H, Deng X, Hu CD (2006) Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis under physiological conditions. Biotechniques 40:61–66
Hudry B, Viala S, Graba Y, Merabet S (2011) Visualization of protein interactions in living Drosophila embryos by the bimolecular fluorescence complementation assay. BMC Biol 9:5
Bischof J, Maeda RK, Hediger M, Karch F, Basler K (2007) An optimized transgenesis system for Drosophila using germ-line-specific phiC31 integrases. Proc Natl Acad Sci U S A 104:3312–3317
Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J et al (2013) A versatile platform for creating a comprehensive UAS-ORFeome library in Drosophila. Development 140: 2434–2442
Gohl C, Banovic D, Grevelhorster A, Bogdan S (2010) WAVE forms hetero- and homo-oligomeric complexes at integrin junctions in Drosophila visualized by bimolecular fluorescence complementation. J Biol Chem 285: 40171–40179
de Navas L, Foronda D, Suzanne M, Sanchez-Herrero E (2006) A simple and efficient method to identify replacements of P-lacZ by P-Gal4 lines allows obtaining Gal4 insertions in the bithorax complex of Drosophila. Mech Dev 123:860–867
Gebelein B, Culi J, Ryoo HD, Zhang W, Mann RS (2002) Specificity of Distalless repression and limb primordia development by abdominal Hox proteins. Dev Cell 3: 487–498
Acknowledgement
This work was supported by the ARC (Association pour la Recherche sur le Cancer), and FRM (Fondation pour la Recherche Médicale).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Duffraisse, M., Hudry, B., Merabet, S. (2014). Bimolecular Fluorescence Complementation (BiFC) in Live Drosophila Embryos. In: Graba, Y., Rezsohazy, R. (eds) Hox Genes. Methods in Molecular Biology, vol 1196. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1242-1_19
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1242-1_19
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1241-4
Online ISBN: 978-1-4939-1242-1
eBook Packages: Springer Protocols