An improved bimolecular fluorescence complementation tool based on   superfolder green fluorescent protein

doi:10.1093/abbs/gmq128

CORC > 北京大学 > 化学与分子工程学院

	An improved bimolecular fluorescence complementation tool based on superfolder green fluorescent protein
	Zhou, Jun ; Lin, Jian ; Zhou, Cuihong ; Deng, Xiaoyan ; Xia, Bin
刊名	acta biochimica et biophysica sinica
	2011
关键词	bimolecular fluorescence complementation protein complementation assays superfolder GFP site-directed gene mutagenesis LIVING CELLS IN-VITRO MAMMALIAN-CELLS VISUALIZATION FRAGMENTS VIVO ASSAYS BIFC
DOI	10.1093/abbs/gmq128
英文摘要	Bimolecular fluorescence complementation (BiFC) has been widely used in the analysis of protein-protein interactions (PPIs) in recent years. There are many notable advantages of BiFC such as convenience and direct visualization of PPI in cells. However, BiFC has one common limitation: the separated non-fluorescent fragments can be spontaneously self-assembled into an intact protein, which leads to false-positive results. In this study, a pair of complementary fragments (sfGFPN and sfGFPC) was constructed by splitting superfolder GFP (sfGFP) between the 214 and 215 amino acid residue, and sfGFPC was mutated by site-directed gene mutagenesis to decrease the signal of negative control. Our results showed that mutations in sfGFPC (sfGFPC(m12)) can effectively decrease the signal of negative control. Thus, we provide an improved BiFC tool for the analysis of PPI. Further, since the self-assembly problem is a common shortcoming for application of BiFC, our research provides a feasible strategy for other BiFC candidate proteins with the same problem.; Biochemistry & Molecular Biology; Biophysics; SCI(E); 中国科技核心期刊(ISTIC); 中国科学引文数据库(CSCD); 8; ARTICLE; 3; 239-244; 43
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/394925]
专题	化学与分子工程学院生命科学学院
推荐引用方式 GB/T 7714	Zhou, Jun,Lin, Jian,Zhou, Cuihong,et al. An improved bimolecular fluorescence complementation tool based on superfolder green fluorescent protein[J]. acta biochimica et biophysica sinica,2011.
APA	Zhou, Jun,Lin, Jian,Zhou, Cuihong,Deng, Xiaoyan,&Xia, Bin.(2011).An improved bimolecular fluorescence complementation tool based on superfolder green fluorescent protein.acta biochimica et biophysica sinica.
MLA	Zhou, Jun,et al."An improved bimolecular fluorescence complementation tool based on superfolder green fluorescent protein".acta biochimica et biophysica sinica (2011).