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. 2017:1668:127-134.
doi: 10.1007/978-1-4939-7283-8_9.

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

Manami Kodaka  1 , Xiaoyin Xu  1   2 , Zeyu Yang  3 , Junichi Maruyama  1 , Yutaka Hata  4   5
Affiliations

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

Manami Kodaka et al. Methods Mol Biol. 2017.

Abstract

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.

Keywords: C2C12 cells; Green fluorescent protein; Myoblast; Myofusion; Myogenesis.

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