Combinational biosynthesis and characterization of fusion proteins with tandem repeats of allophycocyanin holo-alpha subunits, and their application as bright fluorescent labels for immunofluorescence assay

doi:10.1016/j.jbiosc.2018.06.004

CORC > 海洋研究所 > 中国科学院海洋研究所 > 实验海洋生物学重点实验室

	Combinational biosynthesis and characterization of fusion proteins with tandem repeats of allophycocyanin holo-alpha subunits, and their application as bright fluorescent labels for immunofluorescence assay
	Chen, Huaxin 1,2,3; Jiang, Peng 1,2,3
刊名	JOURNAL OF BIOSCIENCE AND BIOENGINEERING
	2018-12-01
卷号	126 期号:6 页码:778-782
关键词	Allophycocyanin Streptavidin Fusion protein Fluorescence Immunofluorescence assay
ISSN号	1389-1723
DOI	10.1016/j.jbiosc.2018.06.004
通讯作者	Chen, Huaxin(chenhx001@126.com)
英文摘要	Fusion protein of streptavidin and allophycocyanin holo-alpha subunit (ApcA) is fluorescent and is able to bind biotin. This fusion protein (SLA) can be used as fluorescent label in immunofluorescence assay. In this study, one or two repeats of ApcA were fused to the protein SLA, with the aim to improve its brightness. The fusion proteins SLA2 (two repeats of ApcA) and SLA3 (three repeats of ApcA), together with lyase (cpcS) and phycoerythrobilin synthesizing enzymes (Ho1 and PebS), were co-expressed in Escherichia coli. These fusion proteins were purified by affinity chromatography. While SLA2 and SLA3 shared similar absorbance spectra, fluorescence spectra and biotin-binding activities with SLA, their brightness were much higher than that of SLA. When used as fluorescent labels in immunofluorescence assay, SLA2 and SLA3 showed higher detection sensitivity than SLA. These results suggested that SLA2 and SLA3 were the preferable fluorescent labels in immunofluorescence assays. (C) 2018, The Society for Biotechnology, Japan. All rights reserved.
资助项目	Development Funds of Science and Technology of Shinan District[2016-3-010-ZH] ; Strategic Priority Research Program of the Chinese Academy of Sciences[XDA11030202] ; National 863 Program[2014AA093505] ; National 863 Program[2014AA093501]
WOS研究方向	Biotechnology & Applied Microbiology ; Food Science & Technology
语种	英语
出版者	SOC BIOSCIENCE BIOENGINEERING JAPAN
WOS记录号	WOS:000452933700017
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/155790]
专题	海洋研究所_实验海洋生物学重点实验室
通讯作者	Chen, Huaxin
作者单位	1.Chinese Acad Sci, Inst Oceanol, CAS Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266071, Peoples R China 3.Chinese Acad Sci, Ctr Ocean Mega Sci, Qingdao 266071, Peoples R China
推荐引用方式 GB/T 7714	Chen, Huaxin,Jiang, Peng. Combinational biosynthesis and characterization of fusion proteins with tandem repeats of allophycocyanin holo-alpha subunits, and their application as bright fluorescent labels for immunofluorescence assay[J]. JOURNAL OF BIOSCIENCE AND BIOENGINEERING,2018,126(6):778-782.
APA	Chen, Huaxin,&Jiang, Peng.(2018).Combinational biosynthesis and characterization of fusion proteins with tandem repeats of allophycocyanin holo-alpha subunits, and their application as bright fluorescent labels for immunofluorescence assay.JOURNAL OF BIOSCIENCE AND BIOENGINEERING,126(6),778-782.
MLA	Chen, Huaxin,et al."Combinational biosynthesis and characterization of fusion proteins with tandem repeats of allophycocyanin holo-alpha subunits, and their application as bright fluorescent labels for immunofluorescence assay".JOURNAL OF BIOSCIENCE AND BIOENGINEERING 126.6(2018):778-782.