Combinational biosynthesis and characterization of a fluorescent 82 beta-phycocyanin of Spirulina platensis

CORC > 烟台海岸带研究所 > 中国科学院烟台海岸带研究所 > 海岸带生物学与生物资源利用重点实验室 > 海岸带生物学与生物资源利用所重点实验室

	Combinational biosynthesis and characterization of a fluorescent 82 beta-phycocyanin of Spirulina platensis
	Guan XiangYu 2; Zhang WeiJie 3; Chi XiaoYuan 3; Lin HanZhi 3,4; Wang JinFeng 3; Qin Song1
刊名	CHINESE SCIENCE BULLETIN
	2012-09-01
卷号	57 期号:25 页码:3294-3299
关键词	Escherichia coli phycocyanin fluorescence recombinant
ISSN号	1001-6538
通讯作者	Qin, S (reprint author), Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China.,sqin@yic.ac.cn
产权排序	[Qin Song] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China; [Guan XiangYu] China Univ Geosci, Sch Ocean Sci, Beijing 100083, Peoples R China; [Zhang WeiJie; Chi XiaoYuan; Lin HanZhi; Wang JinFeng] Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China; [Lin HanZhi] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
中文摘要	To biosynthesize fluorescent Spirulina platensis (Sp) beta-phycocyanin (PC) in Escherichia coli, a BLASTP search for homologs of the cpeS gene, a chromophore lyase, was performed against the Synechocystis sp. PCC 6803 (S6) proteome. A highly homologous gene, slr2049, was obtained from the S6 genome. Sites 82 and 153 in beta-phycocyanin of Sp were modified by site-directed mutagenesis. Two recombinant expression vectors were constructed and transformed into E. coli BL21: (i) pCDF-cpcB (C153A)-slr2049-sll0583-ho1-pcyA; and (ii) pCDF-cpcB (C82I)-slr2049-sll0583-ho1-pcyA. Lyases encoded by the genes slr2049 and sll0583 catalyzed the linking of Sp 82 beta-PC to phycocyanobilin (PCB), and fluorescent CpcB (C153A)-PCB was generated. We present a strategy for the co-expression of multiple genes in a single expression vector to identify the function of an unknown gene. Recombinant phycobiliproteins produced on a large scale are promising fluorescent tags for diagnostics and pharmacology.
英文摘要	To biosynthesize fluorescent Spirulina platensis (Sp) beta-phycocyanin (PC) in Escherichia coli, a BLASTP search for homologs of the cpeS gene, a chromophore lyase, was performed against the Synechocystis sp. PCC 6803 (S6) proteome. A highly homologous gene, slr2049, was obtained from the S6 genome. Sites 82 and 153 in beta-phycocyanin of Sp were modified by site-directed mutagenesis. Two recombinant expression vectors were constructed and transformed into E. coli BL21: (i) pCDF-cpcB (C153A)-slr2049-sll0583-ho1-pcyA; and (ii) pCDF-cpcB (C82I)-slr2049-sll0583-ho1-pcyA. Lyases encoded by the genes slr2049 and sll0583 catalyzed the linking of Sp 82 beta-PC to phycocyanobilin (PCB), and fluorescent CpcB (C153A)-PCB was generated. We present a strategy for the co-expression of multiple genes in a single expression vector to identify the function of an unknown gene. Recombinant phycobiliproteins produced on a large scale are promising fluorescent tags for diagnostics and pharmacology.
学科主题	Multidisciplinary Sciences
研究领域[WOS]	Science & Technology - Other Topics
关键词[WOS]	SYNECHOCOCCUS SP PCC-7002 ; BLUE-GREEN-ALGAE ; C-PHYCOCYANIN ; ESCHERICHIA-COLI ; ALPHA-PHYCOCYANIN ; FERREDOXIN OXIDOREDUCTASE ; CHROMOPHORE ATTACHMENT ; ANTIOXIDANT ACTIVITY ; EXPRESSION ; LYASE
收录类别	SCI
资助信息	Jiangsu Provincial Marine Biotechnology Key Lab Open Foundation[2010HS02]; National Natural Science Foundation of China[41006094]
语种	英语
WOS记录号	WOS:000307750500007
公开日期	2013-03-08
内容类型	期刊论文
源URL	[http://ir.yic.ac.cn/handle/133337/6097]
专题	烟台海岸带研究所_海岸带生物学与生物资源利用所重点实验室
作者单位	1.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China 2.China Univ Geosci, Sch Ocean Sci, Beijing 100083, Peoples R China 3.Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China 4.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Guan XiangYu,Zhang WeiJie,Chi XiaoYuan,et al. Combinational biosynthesis and characterization of a fluorescent 82 beta-phycocyanin of Spirulina platensis[J]. CHINESE SCIENCE BULLETIN,2012,57(25):3294-3299.
APA	Guan XiangYu,Zhang WeiJie,Chi XiaoYuan,Lin HanZhi,Wang JinFeng,&Qin Song.(2012).Combinational biosynthesis and characterization of a fluorescent 82 beta-phycocyanin of Spirulina platensis.CHINESE SCIENCE BULLETIN,57(25),3294-3299.
MLA	Guan XiangYu,et al."Combinational biosynthesis and characterization of a fluorescent 82 beta-phycocyanin of Spirulina platensis".CHINESE SCIENCE BULLETIN 57.25(2012):3294-3299.