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NCUEIR > College of Science > Department of Biology > Periodical Articles > Item 987654321/16767

Please use this identifier to cite or link to this item: http://ir.ncue.edu.tw/ir/handle/987654321/16767

Title:	Molecular Cloning of a cDNA Coding for Copper/Zinc Superoxide Dismutase from Zebrafish and Its Expression in Escherichia coli
Authors:	Ken, Chuian-Fu;Shaw, Jei-Fu;Wu, Jen-Leih;Lin, Chi-Tsai
Contributors:	生物學系
Keywords:	cDNA;Copper / zinc superoxide dismutase;Danio rerio;Escherichia coli;Expression;Molecular cloning;PCR;pET-23a(+)-thioredoxin;Zebrafish
Date:	1998-07
Issue Date:	2013-06-05T08:48:06Z
Publisher:	American Chemical Society
Abstract:	A full-length complementary DNA (cDNA) clone encoding a putative copper/zinc Superoxide dismutase (Cu/Zn-SOD) was amplified by a polymerase chain reaction (PCR)-based technique from cDNA synthesized from zebrafish (Danio rerio) mRNA. Nucleotide sequence analysis of this cDNA clone revealed that it comprised a complete open reading frame coding for 154 amino acid residues. The deduced amino acid sequence showed higher identity (73.5-74.3%) with swordfish and shark Cu/Zn-SOD than with Cu/Zn-SOD from mammals (69.6-70.9%) and plants (55.8-56.2%). The ammo acid residues required for coordinating copper and zinc are conserved, as they are present in all reported Cu/Zn-SOD sequences. It lacks a targeting sequence, which suggests that the zebrafish cDNA clone encodes a cytosolic Cu/Zn-SOD. Furthermore, the coding region of Cu/Zn-SOD from zebrafish was introduced into an expression vector, pET-23a(+)-thioredoxin, and transformed into Escherichia coli AD494(DE3)pLysS. A predominant achromatic zone was detected by activity staining of native PAGE, and the expression pattern was shown by Coomassie blue staining of SDS-PAGE. This indicates that the Cu/Zn-SOD cDNA clone can be expressed in E. coli.
Relation:	Journal of Agricultural and Food Chemistry, 46(7): 2863-2867
Appears in Collections:	[Department of Biology] Periodical Articles

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