Protein-protein interactions in Drosophila myofibrils are essential for their function
and formation. Bimolecular Fluorescence Complementation (BiFC) is an effective method
for studying protein interactions and localization. BiFC relies on the reconstitution
of a monomeric fluorescent protein from two half-fragments when in proximity. Two
proteins tagged with the different half-fragments emit a fluorescent signal when they
are in physical contact, thus revealing a protein interaction and its spatial distribution.
Because myofibrils are large networks of interconnected proteins, BIFC is an ideal
method to study protein-protein interactions in myofibrils. Here we present a protocol
for generating transgenic flies compatible with BiFC and a method for analyzing protein-protein
interactions based on the fluorescent BiFC signal in myofibrils. Our protocol is applicable
to the majority of Drosophila proteins and with few modifications may be used to study
any tissue.
