PUBLICATION
Molecular cloning of a cDNA coding for copper/zinc superoxide dismutase from zebrafish and its expression in Escherichia coli
- Authors
- Ken, C.-F., Shaw, J.-F., Wu, J.-L., and Lin, C.-T.
- ID
- ZDB-PUB-980810-29
- Date
- 1998
- Source
- Journal of Agricultural and Food Chemistry 46: 2863-2867 (Journal)
- Registered Authors
- Shaw, Jei-Fu
- Keywords
- zebrafish; Danio rerio; molecular cloning; cDNA; copper/zinc superoxide dismutase; expression; Escherichia coli; PCR; pET-23a(+)-thioedoxin
- MeSH Terms
- none
- PubMed
- none Full text @ J. Agric. Food Chem.
Citation
Ken, C.-F., Shaw, J.-F., Wu, J.-L., and Lin, C.-T. (1998) Molecular cloning of a cDNA coding for copper/zinc superoxide dismutase from zebrafish and its expression in Escherichia coli. Journal of Agricultural and Food Chemistry. 46:2863-2867.
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 (73.574.3%) with swordfish and shark Cu/Zn-SOD than with Cu/Zn-SOD from mammals (69.670.9%) and plants (55.856.2%). The amino 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 SDSPAGE. This indicates that the Cu/Zn-SOD cDNA clone can be expressed in E. coli.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping