利用分裂绿色荧光蛋白重组检测法体外研究成肌作用和肌融合

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

作者信息

Kodaka Manami, Xu Xiaoyin, Yang Zeyu, Maruyama Junichi, Hata Yutaka

机构信息

Department of Medical Biochemistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan.

Department of Breast Surgery, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.

出版信息

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

DOI:10.1007/978-1-4939-7283-8_9

PMID:28842906

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.

摘要

绿色荧光蛋白（GFP）由11条β链组成，当被分为N端的十条β链（GFP1-10）和C端的最后一条β链（GFP11）时，会失去GFP信号。然而，当GFP1-10和GFP11相遇时，它们会重新组装成有荧光的GFP。我们在C2C12细胞中表达了GFP1-10和与杀稻瘟菌素抗性基因产物融合的GFP11（BSR-GFP11）。这两种细胞系均不显示GFP，但当它们进行肌生成和肌融合时，GFP1-10和BSR-GFP11在多核肌管中形成荧光复合物，从而使GFP信号反映肌生成和肌融合。