title: Metal-binding activity of the soluble recombinant pig metallothionein 1A expressed in Escherichia coli.
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⁺).
authors: College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing High-tech Industrial Development Zone, Daqing, 163319, People's Republic of China. dongbosun@yahoo.com.cn
authors: Dongbo
authors: D
authors: Sun
authors: Hong
authors: H
authors: Zhang
authors: Guojun
authors: G
authors: Wu
authors: Qinghe
authors: Q
authors: Zhu
authors: Siwen
authors: S
authors: Lv
authors: Donghua
authors: D
authors: Guo
authors: Rui
authors: R
authors: Wu
authors: Jun
authors: J
authors: Bao
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002104:Cadmium / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002614:Chelating Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D003062:Codon; D003300:Copper / Q000378:metabolism; D003956:Dialysis; D007700:Kinetics; D008668:Metallothionein / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008958:Models, Molecular; D008970:Molecular Weight; D011487:Protein Conformation; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D034421:Sus scrofa; D015032:Zinc / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D002614:Chelating Agents; D003062:Codon; D020033:Protein Isoforms; D011994:Recombinant Proteins; D002104:Cadmium; D003300:Copper; D008668:Metallothionein; D015032:Zinc