Abstract：
:The bimolecular fluorescence complementation (BiFC) assay is a powerful tool for visualizing and identifying protein interactions in living cells. This assay is based on the principle of protein-fragment complementation, using two nonfluorescent fragments derived from fluorescent proteins. When two fragments are brought together in living cells by tethering each to one of a pair of interacting proteins, fluorescence is restored. Here, we provide a protocol for a Venus-based BiFC assay to visualize protein interactions in the living nematode, Caenorhabditis elegans. We discuss how to design appropriate C. elegans BiFC cloning vectors to enable visualization of protein interactions using either inducible heat shock promoters or native promoters; transform the constructs into worms by microinjection; and analyze and interpret the resulting data. When expression of BiFC fusion proteins is induced by heat shock, the fluorescent signals can be visualized as early as 30 min after induction and last for 24 h in transgenic animals. The entire procedure takes 2-3 weeks to complete.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Shyu YJ,Hiatt SM,Duren HM,Ellis RE,Kerppola TK,Hu CDdoi
10.1038/nprot.2008.16subject
Has Abstractpub_date
2008-01-01 00:00:00pages
588-96issue
4eissn
1754-2189issn
1750-2799pii
nprot.2008.16journal_volume
3pub_type
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