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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, Huaxin1,2,3; Jiang, Peng1,2,3
2018-12-01
发表期刊JOURNAL OF BIOSCIENCE AND BIOENGINEERING
ISSN1389-1723
卷号126期号:6页码:778-782
通讯作者Chen, Huaxin(chenhx001@126.com)
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.
关键词Allophycocyanin Streptavidin Fusion protein Fluorescence Immunofluorescence assay
DOI10.1016/j.jbiosc.2018.06.004
收录类别SCI
语种英语
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] ; 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
WOS类目Biotechnology & Applied Microbiology ; Food Science & Technology
WOS记录号WOS:000452933700017
出版者SOC BIOSCIENCE BIOENGINEERING JAPAN
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被引频次:7[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/155790
专题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
第一作者单位中国科学院海洋研究所
通讯作者单位中国科学院海洋研究所
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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.
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