A plant metallothionein produced in E. coli

FEBS Letters

Winge, Dennis R.²  Harwood, John L.¹  Kay, John¹  Kille, Peter¹ 

[1] Department of Biochemistry, University of Wales College of Cardiff, Cardiff CF1 1ST, Wales, UK;Haematology/Oncology Division, University of Utah School of Medicine, Salt Lake City, UT 84132, USA

关键词: Pea metallothionein;    Cloning;    PCR;    Expression in E. coli;    Recombinant metallothionein;    Extended linker region;    Proteolytic nicking;    Cysteine-rich region;    Metal-binding;   

DOI  :  10.1016/0014-5793(91)81411-Z

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A metallothionein cDNA was generated from pea (Pisum sativum L.) roots, amplified by PCR and inserted into a plasmid for expression in E. coli. Purification of the resultant product generated 3 pools of cadmium-containing material after DEAE-cellulose chromatography. The amino acid composition of each was in excellent agreement with that predicted for pea metallothionein. A cadmium content of ∼6 g.atoms per mole of protein was estimated. N-terminal sequence analysis revealed that the recombinant molecule had been proteolysed within the extended region linking the 2 cysteine-rich (putative) metal-binding regions. The significance of these findings in terms of the protein folding/targeting of the molecule are considered.

【 授权许可】

Unknown