Abstract
Full-length cDNA for the pig metallothionein 1A (pMT1A) gene was synthesized based on the pig MT1A gene sequence in Genbank and cloned into the pMD18-T vector. After sequence analysis and structure prediction, the pMT1A gene was cloned into vector pET-32a (+) containing a His-tag. The recombinant pMT1A (rpMT1A) was expressed in a soluble form using Escherichia coli Rosetta™ (DE3) plysS cells. Western blotting showed that the purified rpMT1A protein bound an anti-His-tag monoclonal antibody. Further investigation revealed that the rpMT1A protein showed high metal-binding activity with the divalent metal ions copper (Cu²⁺), zinc (Zn²⁺), and cadmium (Cd²⁺).
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Blotting, Western
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Cadmium / metabolism
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Carrier Proteins / chemistry
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Carrier Proteins / genetics
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Carrier Proteins / isolation & purification
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Carrier Proteins / metabolism*
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Chelating Agents / chemistry
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Chelating Agents / isolation & purification
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Chelating Agents / metabolism*
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Codon
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Copper / metabolism
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Dialysis
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Kinetics
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Metallothionein / chemistry
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Metallothionein / genetics
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Metallothionein / isolation & purification
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Metallothionein / metabolism*
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Models, Molecular
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Molecular Weight
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Protein Conformation
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Protein Isoforms / chemistry
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Protein Isoforms / genetics
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Protein Isoforms / isolation & purification
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Protein Isoforms / metabolism
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Solubility
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Sus scrofa
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Zinc / metabolism
Substances
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Carrier Proteins
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Chelating Agents
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Codon
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Protein Isoforms
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Recombinant Proteins
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Cadmium
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Copper
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Metallothionein
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Zinc