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標題: Cloning and expression of a seed-specific metallothionein-like protein from sesame
作者: Chyan, C.L.
Lee, T.T.T.
Liu, C.P.
Yang, Y.C.
Tzen, J.T.C.
Chou, W.M.
關鍵字: expression;GST-fusion;metallothionein-like;seed;sesame;cdna clones;gene;zinc;copper;embryogenesis;arabidopsis;superoxide;storage;stress;yeast
Project: Bioscience Biotechnology and Biochemistry
期刊/報告no:: Bioscience Biotechnology and Biochemistry, Volume 69, Issue 12, Page(s) 2319-2325.
A cDNA clone, SiMT encoding an Ec type of metallothionein (MT)-like protein, was isolated from maturing seeds of sesame (Sesamum indicum L.), and its deduced protein sequence shared 47-65% similarity to other known Ec type of MT-like proteins with three highly conserved cysteine-rich segments. The transcript of SiMT was exclusively accumulated in maturing seeds from two weeks after flowering to the end of seed maturation. The results of a southern blot analysis suggested that one SiMT and one SiMT-like gene were present in the sesame genome. Recombinant SiMT fused with glutathione-S-transferase (GST) was over-expressed in Escherichia coli, and purified to homogeneity by affinity chromatography. Recombinant SiMT released from GST was harvested after factor Xa cleavage. Migration of the recombinant SiMT during SDS-PAGE was accelerated when its binding metal ions were depleted by EDTA. The metal-binding capability of recombinant SiMT was measured by inductively-coupled plasma atomic emission spectrometry. Our results show that the recombinant SiMT could trap zinc or copper ions, but not manganese ions, with a stoichiometric ratio (metal ion/SiMT) of approximately 2.
ISSN: 0916-8451
DOI: 10.1271/bbb.69.2319
Appears in Collections:生物科技學研究所

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