Overexpression of petH gene of Cyanobacterium synechococcus sp. PCC 7002   in Escherichia coli and purification of the expressed product

CORC > 北京大学 > 生命科学学院

	Overexpression of petH gene of Cyanobacterium synechococcus sp. PCC 7002 in Escherichia coli and purification of the expressed product
	Li, RG ; Zhao, JD ; Wu, GY ; Wu, XY
刊名	植物学报
	1998
关键词	petH gene FNR electron transport overexpression protein refolding FERREDOXIN-NADP+ OXIDOREDUCTASE REDUCTASE
英文摘要	Fd:NADP(+) oxidoreductase (FNR) is one of the key enzymes in photosynthetic electron transport. The gene petH encoding FNR of Synechococcus sp. PCC 7002 was cloned into the expressing vector pET-3 d' and overexpressed in E, coli. The amount of recombinant FNR (rFNR) was over 50% of the total cellular proteins. There were two forms of FNR activity, one is soluble and the other one was in the form of inclusion bodies. The soluble rFNR was purified through ion exchange chromatography and gel chromatography. The rFNR in the form of inclusion bodies was first solubilized with 6.7 mol/L urea, and then refolded into the active form in the presence of flavin adenine dinucleotide (FAD). Further purification was performed by ion exchange chromatography. The rFNR pruified from either form of the expressed product had the maximum absorption spectrum as that of the natural FNR from cyanobacteria, whose maximum absorption was at 273, 385 and 456 nm respectively. N-terminal sequencing showed that rFNR was indeed a product of petH gene expression. rFNR could catalyze the electron transport from P-700 to NADP(+) in the presence of ferredoxin. The optimal pH for diaphorase activity of rFNR was 8.0 and the optimal temperature was 30 degrees C.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000076577300007&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Biochemistry & Molecular Biology; Plant Sciences; SCI(E); 中文核心期刊要目总览(PKU); 中国科学引文数据库(CSCD); 0; 9; 814-819; 40
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/390074]
专题	生命科学学院
推荐引用方式 GB/T 7714	Li, RG,Zhao, JD,Wu, GY,et al. Overexpression of petH gene of Cyanobacterium synechococcus sp. PCC 7002 in Escherichia coli and purification of the expressed product[J]. 植物学报,1998.
APA	Li, RG,Zhao, JD,Wu, GY,&Wu, XY.(1998).Overexpression of petH gene of Cyanobacterium synechococcus sp. PCC 7002 in Escherichia coli and purification of the expressed product.植物学报.
MLA	Li, RG,et al."Overexpression of petH gene of Cyanobacterium synechococcus sp. PCC 7002 in Escherichia coli and purification of the expressed product".植物学报 (1998).