Cloning, expression and characterization of metallothionein from Antarctic clam Laternula elliptica
- Cloning, expression and characterization of metallothionein from Antarctic clam Laternula elliptica
- Other Titles
- 남극큰띠조개로부터 metallothionein 클로닝
- Ahn, In-Young
- Antarctic; Laternula elliptica; metallothioneins
- Issue Date
- Ahn, In-Young, et al. 2006. Cloning, expression and characterization of metallothionein from Antarctic clam Laternula elliptica. 2006.05.09~.
- The genes for two apparent subtypes of metallothioneins (MTs) isoform were isolated from Antarctic clam, Laternula elliptica.
Determination of the nucleotide sequence showed that the gene consist of 222 bp that codes a 73-amino acid protein. The
comparison between MTs cDNA sequences of L. elliptica and other bivalves shows strong homologies on positions of cysteine
residues which are to be important for their metal binding abilities. The gene for the metallothionein was inserted into a pET vector
and overexpressed as carboxyl terminal extension of glutathionein-S-transferase (GST) in Escherichia coli. After the GST fusion
proteins had been purified by glutathione-Sepharose affinity chromatography column and digested with enterokinase, the
metallothionein was purified with gel filtration and analyzed for their biochemical properties. Recombinant MTs were reconstituted
with Cd, Cu, and Zn, and kinetic studies of the reactions with electrophilic disulphide, DTNB, were investigated to explore the metal
binding ability. It is reveal that the Cd-MT and Zn-Mt react with DTNB biphasically and Zn-MT reacts with DTNB more rapidly and
significantly greater observed pseudo-first order rate constant. Cu-MT react monophasic and release metal slowly from
- Conference Date
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