Application of Split-GFP Reassembly Assay to Study Myogenesis and Myofusion In Vitro

Green fluorescent protein (GFP) is composed of 11 β-strands, and loses GFP signals, when divided into the N-terminal ten β-strands (GFP1–10) and the C-terminal last β-strand (GFP11). However, when GFP1–10 and GFP11 encounter, they reassemble into the fluorescent GFP. We expressed GFP1–10 and blasticidin resistance gene product-fused GFP11 (BSR-GFP11) in C2C12 cells. Both the cell lines do not show GFP but when they undergo myogenesis and myofusion, GFP1–10 and BSR-GFP11 form the fluorescent complex in multi-nuclear myotubes, so that GFP signals reflect myogenesis and myofusion.